TW200538545A - Method of manufacturing liquid koji - Google Patents

Method of manufacturing liquid koji Download PDF

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Publication number
TW200538545A
TW200538545A TW094110948A TW94110948A TW200538545A TW 200538545 A TW200538545 A TW 200538545A TW 094110948 A TW094110948 A TW 094110948A TW 94110948 A TW94110948 A TW 94110948A TW 200538545 A TW200538545 A TW 200538545A
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TW
Taiwan
Prior art keywords
liquid
rice
culture
shochu
brewing
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TW094110948A
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Chinese (zh)
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TWI347360B (en
Inventor
Toshikazu Sugimoto
Hiroshi Kouji
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Asahi Breweries Ltd
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Priority claimed from JP2004350661A external-priority patent/JP3718677B2/en
Priority claimed from JP2004352320A external-priority patent/JP3718678B1/en
Priority claimed from JP2004352324A external-priority patent/JP3718679B1/en
Priority claimed from JP2004378453A external-priority patent/JP3718681B1/en
Application filed by Asahi Breweries Ltd filed Critical Asahi Breweries Ltd
Publication of TW200538545A publication Critical patent/TW200538545A/en
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Publication of TWI347360B publication Critical patent/TWI347360B/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12H—PASTEURISATION, STERILISATION, PRESERVATION, PURIFICATION, CLARIFICATION OR AGEING OF ALCOHOLIC BEVERAGES; METHODS FOR ALTERING THE ALCOHOL CONTENT OF FERMENTED SOLUTIONS OR ALCOHOLIC BEVERAGES
    • C12H6/00—Methods for increasing the alcohol content of fermented solutions or alcoholic beverages
    • C12H6/02—Methods for increasing the alcohol content of fermented solutions or alcoholic beverages by distillation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
    • C12G3/00—Preparation of other alcoholic beverages
    • C12G3/02—Preparation of other alcoholic beverages by fermentation
    • C12G3/021—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn
    • C12G3/022—Preparation of other alcoholic beverages by fermentation of botanical family Poaceae, e.g. wheat, millet, sorghum, barley, rye, or corn of botanical genus Oryza, e.g. rice

Abstract

An object of the present invention is to provide a liquid koji to be used in the production of a fermented food or drink, particularly a liquid koji having glucoamylase and acid-stable α -amylase with high enzymatic activities, which can be used in the brewing of shochu. The present invention provides a method of manufacturing a liquid koji to be used inproduction of a fermented food or drink, comprising: cultivating a koji mold in a liquid medium that contains, as a raw material, a substance selected from the group consisting of a cereal having a surface covered with a husk; a cereal having a surface from which only a husk (e.g., a chaff) is removed; an unprocessed bean or tuber having a surface covered with a hull; and amaranthus and/or quinoa. According to the present invention, both enzymes of glucoamylase and acid-stable α -amylase are simultaneously produced with high yield in a balanced manner to allow the production of a liquid koji having enzymatic activities required for, for example, the brewing of shochu. By using the liquid koji, fermented foods and drinks such as shochu can be efficiently produced.

Description

200538545 九、發明說明： 【發明所屬之技術領域】 本發明係相關於製造發酵飲食品時使用的液體麴，特別 是具備釀造燒酒時所需的酵素活性的液體麴之製法。 【先前技術】 製造酒類時使用的趨有將絲狀菌的抱子接種於經蒸煮 . 等處理後的原料而培養之固體麴，以及將原料和其他營養 源添加於水調製成液體培養基，再接種麴菌的孢子或前培 養的菌絲等進行培養之液體麴。 B 先前的酒類或發酵飲食品例如日本清酒、燒酒、醬油、 味噌、味酣等的製造過程中，廣泛使用固體培養法進行製 麴，亦即採用固體麴。此固體培養法係將白麴菌 (Aspergillus kawachii)、泡盛麴菌（Aspergillus awamori)、 黑麴菌（Aspergillus niger)、黃麴菌（Aspergillus oryzae)、 或醬油麴菌（Aspergillus sojae)等麴菌的孢子散佈於已蒸 煮的榖類等固體原料，使麹菌在表面增殖之培養方法。 例如製造燒酒時，廣泛使用白麹菌（Aspergillus φ kawachii)、泡盛麴菌（Aspergillus awamori)等固體麴。 惟，固體培養法係一種原料或麴菌不均勻分散的培養系， 不易使溫度或水分含量、各種營養成分等因素一致，控制 其培養的機制相當複雜。又，多在開放狀態下製麴，此時 易受雜菌污染在品質管理方面需多注意。因此，此法不適 用於大規模的製造。 相對於此，液體培養法可容易地進行培養控制或品質管 理，係一種可有效地生產之培養型態，例如，因無法充分 200538545 具有釀造燒酒所需的酵素活性時，極少 養所得的培養物作爲燒酒麴。在此，由 培養物，除了指從液體培養法收得的培 麴）之外，亦指培養液、菌體、其濃縮 由液體培養法製得的培養物無法使 食品的製造之主要原因，係液體培養時 維素酶等酵素生成機制大不同於固體培 成性較低落（參考非專利文獻1 )。 通常，以燒酒爲首的酒類之製造，以 酒精。因此，影響饋給麴菌的葡萄糖之 關酵素，特別是葡糖丨殿粉酶或耐酸性α 酵過程之關鍵酵素。惟，由液體培養法 葡糖澱粉酶的活性顯著降低，其生成機 不相同（參考非專利文獻2 )。 提昇麴菌的葡糖澱粉酶的活性之方注 的階段施予壓力處理之同時培養麴菌之 獻1 )或在麴菌培養液中添加焙炒的穀 利文獻2)等。專利文獻1中的方法， 性膜或空隙的固定化劑中，發現標識葡 因glaB而提高同酵素活性，需要嚴密的 裝置，較不實用。又，專利文獻2中的 養於使用至少部分的原料經過焙炒的穀 焙炒穀類亦需增加新穎的製造工程。 本發明者們發明一種使用含有麴菌 液體培養基之麴菌培養方法（參考專利 可將麴菌經液體培 液體培養法收得的 養物（又稱爲液體 物或其乾燥物等。 用於燒酒等發酵飲 麴菌的澱粉酶、纖 養，一般而言其生 並行複發酵使生成 麴菌的糖質分解相 -澱粉酶，係酒精發 製得的培養物中， 制和固體培養法大 丨，例如在菌絲生育 方法（參考專利文 類之方法（參考專 係培養於具有多孔 糖澱粉酶的新穎基 控制及特殊的培養 方法，係將麴菌培 類之液體培養基， 難以分解的糖質的 文獻3 )。根據本 200538545 發明，可簡便且經濟地製得在麴菌的液體培養中，一種可 用於酒類或發酵飮食品的製造的具高活性葡糖澱粉酶等糖 質分解相關酵素之麴菌培養物。 另一方面，最近開始詳細地進行有關耐酸性α -澱粉酶 之分子生物學的解析（參考非專利文獻3)。依據上述的 分析，白麴菌具有非耐酸性α -澱粉酶和耐酸性α -澱粉酶 ▲ 等性質相異的2種類澱粉酶基因，惟其發現樣式大不相 同，液體培養中，可充分生成非耐酸性α-澱粉酶，而幾乎 Β 無法生成釀造燒酒時的關鍵酵素亦即耐酸性α -澱粉酶。 製造燒酒時，爲要預防燒酒酒醴的腐壞，於低pH環境 下進行釀造。惟，因非耐酸性α ·澱粉酶於低PH條件下快 速地失去活性，對釀造燒酒時的糖質分解幾乎無貢獻。因 此’爲要製造燒酒，必須在麴菌的液體培養中大量生成可 分解燒酒釀造時的糖質之耐酸性α -澱粉酶。 先前，亦有探討麴菌的液體培養中，耐酸性α -澱粉酶 的生成機制之報告，其方法係使用含有蛋白腺或檸檬酸緩 衝液的合成培養基，培養時間超過1 00小時等，不適用於 # 實際的燒酒釀造之液體麴製法（參考非專利文獻4)。 專利文獻1 :特開平1 1 - 2 2 5 7 4 6號公報 專利文獻2:特開200 1 -32 1 1 54號公報 專利文獻3 :特開2003-265 1 65號公報 非專利文獻 1 : Iwashita K. et al: Biosci. Biotechnol. Biochem·，62，1938-1946 ( 1998)、山根雄一等：日本釀造 協會誌·，99,84-92 ( 2004 ) 非專利文獻 2: Hata Y. et al: J. Ferment· Bioeng.，84, 200538545 5 3 2-5 3 7( 1 997 )、Hata Y. et al: Gene·，207，1 27- 1 34( 1 99 8 )、200538545 IX. Description of the invention: [Technical field to which the invention belongs] The present invention relates to a method for producing liquid mash used in the production of fermented foods and drinks, in particular, a liquid mash having the enzyme activity required for brewing shochu. [Prior technology] When manufacturing alcoholic beverages, the tassel of filamentous fungi is inoculated into solid mash cultured after cooking and other processed raw materials, and raw materials and other nutrient sources are added to water to prepare a liquid culture medium, and then Inoculated with liquid maggots for spores or pre-cultured hyphae. B. In the previous manufacturing process of alcoholic beverages or fermented food and beverages such as Japanese sake, shochu, soy sauce, miso, miso, etc., solid culture was widely used for making ravioli, that is, solid ravioli. This solid culture method uses Aspergillus kawachii, Aspergillus awamori, Aspergillus niger, Aspergillus oryzae, or Aspergillus sojae. A culture method in which spores are spread on solid raw materials such as boiled maggots to multiply maggots on the surface. For example, in the production of shochu, solid maggots such as Aspergillus φ kawachii and Aspergillus awamori are widely used. However, the solid culture method is a kind of raw material or a heterogeneous culture system in which the fungi are not uniformly dispersed. It is not easy to make factors such as temperature or moisture content, various nutrients, and the control mechanism of the culture is quite complicated. Also, pupae are mostly made in an open state. At this time, susceptibility to contamination by germs should be paid more attention to in terms of quality management. Therefore, this method is not suitable for large-scale manufacturing. In contrast, the liquid culture method can easily perform culture control or quality control, and is a type of culture that can be efficiently produced. For example, when the enzyme activity required for brewing shochu is not sufficient, 200538545, the culture obtained is rarely raised. As shochu ravioli. Here, the term "culture" refers to culture fluids, bacterial cells, and their concentrated cultures produced by the liquid culture method, in addition to the cultivars obtained from the liquid culture method. In liquid culture, enzymes such as vitamine enzymes differ greatly from solid cultures in their low productivity (see Non-Patent Document 1). Generally, alcoholic beverages are produced by shochu. Therefore, the relevant enzymes affecting the glucose fed to the fungi, especially glucose, enzymes or key enzymes that are resistant to acid alpha enzymes. However, the glucoamylase activity is significantly reduced by the liquid culture method, and its production mechanism is different (see Non-Patent Document 2). The method of increasing the glucoamylase activity of Pleurotus ostreatus is to apply pressure treatment at the stage of culturing Pleurotus spp. While cultivating Pleurotus spp. 1) or to add roasted gluten to the Pleurotus spp. Culture solution 2). In the method of Patent Document 1, it is found that the labeling agent increases the activity of the same enzyme due to glaB in the immobilizing agent of the sex film or the void, and requires a strict device, which is not practical. In addition, in Patent Document 2, roasted cereals that are roasted using at least part of the raw materials also require new manufacturing processes. The inventors have invented a method for cultivating maggots using a liquid medium containing maggots (refer to the patent, which can be obtained by liquid culture of maggots through liquid culture (also known as liquid or its dried matter). Used for shochu The amylase and fibrous nutrition of fermented drinking fungi are generally re-fermented in parallel to make the saccharide-decomposing phase of the fungus-amylase, which is produced by alcohol-based culture. For example, in the mycelium growth method (refer to the patent method) (refer to the special cultivation method of the novel base control and special cultivation methods with porous glucoamylase), which is a liquid culture medium of mycelium, which is difficult to decompose. Document 3). According to the invention of 200538545, it is possible to easily and economically prepare a glycolysis-related enzyme having high activity, such as glucoamylase, which can be used in the production of alcoholic beverages or fermented glutinous foods in liquid culture of tadpoles. On the other hand, the molecular biology analysis of acid-resistant α-amylase has recently been carried out in detail (refer to Non-Patent Document 3). Analysis, the white fungus has two types of amylase genes with different properties such as non-acid-resistant α-amylase and acid-resistant α-amylase ▲, but it is found that the patterns are very different. In liquid culture, non-acid-resistant α -Amylase, and almost B can not produce acid-resistant α-amylase, the key enzyme when brewing shochu. When making shochu, it is brewed in a low pH environment in order to prevent the shochu shochu from spoiling. Acid-resistant α · amylase rapidly loses its activity under low pH conditions, and has almost no contribution to the decomposition of sugar during brewing shochu. Therefore, 'to produce shochu, a large amount of decomposable shochu must be produced in the liquid culture of shochu Acid-resistant α-amylase of sugar. Previously, there have been reports on the mechanism of acid-resistant α-amylase production in liquid culture of P. utilis. The method is to use a synthetic medium containing protein glands or citrate buffer. , The culture time is more than 100 hours, etc., it is not suitable for # actual shochu brewing method (refer to Non-Patent Document 4). Patent Document 1: Japanese Patent Application Laid-Open No. 1 1-2 2 5 7 4 6 Patent Document 2: JP 200 1 -32 1 1 54 Patent Document 3: JP 2003-265 1 65 Non-Patent Document 1: Iwashita K. et al: Biosci. Biotechnol. Biochem., 62, 1938- 1946 (1998), Yamane Yuichi, etc .: Japan Brewing Association Journal, 99,84-92 (2004) Non-Patent Document 2: Hata Y. et al: J. Ferment · Bioeng., 84, 200538545 5 3 2-5 3 7 (1 997), Hata Y. et al: Gene ·, 207, 1 27- 1 34 (1 99 8),

Ishita H. e t al: J. Ferment. Bioeng.,8 6, 301-307 ( 1998)、Ishita H. e t al: J. Ferment. Bioeng., 8, 6, 301-307 (1998),

Ishita H. e t al: Curr Genet., 37, 3 7 3 - 3 79 ( 2000 ) 非專利文獻 3: Nagamine K· et al: Biosci. Biotechnol.Ishita H. e t al: Curr Genet., 37, 3 7 3-3 79 (2000) Non-Patent Document 3: Nagamine K. et al: Biosci. Biotechnol.

Biochem.,67, 2 1 94-2202 ( 2003 ) 非專利文獻 4· Sudo S. et al: J. Fernient. Bioeng.，76, 1 05- 1 1 0 ( 1 993 ) ^ Sudo S. et al: J. Ferment. Bioeng.,77, 48 3 -489( 1 994 )、須藤茂俊等··日本釀造協會誌.，8 9,768 ·774 (1 994 ) 【發明內容】 〔解決發明之課題〕 惟，專利文獻3的方法中，具高活性的葡糖澱粉酶之麴 菌培養物，係將麴菌培養於添加難分解性糖質調製成的液 體培養基所生成者，無法使用以榖類等培養原料調製成普 通的液體培養基進行培養。 又，揭示將麴菌培養在液體培養基，製得具高活性的葡 糖澱粉酶的麹菌培養物之技術，惟無提及在液體培養基中 培養麴菌，製得具有高活性的酒精發酵中另一關鍵酵素亦 即耐酸性α -澱粉酶的液體麴之技術。一般認爲，此耐酸性 α -澱粉酶係一種無法於液體培養中生成之酵素，截至目前 爲止’尙未開發出具高活性的耐酸性α -澱粉酶之液體麴。 本發明的目的係提供一種液體麴之製法，其特徵係將製 造發酵飲食品時使用的液體麴，特別是釀造燒酒等酒精發 酵的關鍵酵素葡糖澱粉酶及耐酸性α -澱粉酶的酵素活性 高之液體麴培養於使用未碾製的穀類，亦即榖皮覆蓋表面 200538545 之榖類或只去除表面的榖皮（粗殻）之榖類、未加工豆類 或芋類、或特定的雜穀類作爲原料之液體培養基，而非培 養於添加特殊糖質等，或使用經焙炒處理的原料之特別液 體培養基。 〔解決課題之方法〕 本發明者等爲要解決上述課題經硏究的結果發現，製造 使用於發酵飲食品的製造之液體麴時，如上述般，將麴菌 培養於含有任一種榖皮覆蓋表面之榖類或只去除表面的榖 φ 皮（粗殻）之榖類、外皮覆蓋在表面的未加工豆類或芋類、 或雜榖類例如寛紫及/或奎藜好（qUin〇a)作爲培養原料之液 體培養基，藉此，可製造葡糖澱粉酶活性、及耐酸性α _殿 粉酶活性已增強之液體麴。又，此酵素活性已增強之液體 麴適用於燒酒的釀造，而完成本發明。 亦即，本發明提供下列所示者。 (1) 一種液體麴之製法，其特徵係用於製造發酵飲食 品時的液體麴之製法，將麹菌培養於含有任一種榖皮覆蓋 在表面之榖類；只去除表面的榖皮（粗殻）之榖類；外皮 ^覆蓋在表面的未加工的豆類或芋類；及莧紫及/或奎藜籽 U u i η 〇 a)作爲培養原料之液體培養基。 (2) 如第1項之液體麹之製法，其中榖皮覆蓋在表面 之榖類係未碾製、或碾製比率爲至少有榖皮殘留在榖粒的 表面者。 (3 )如第1或2項之液體麹之製法，其中榖類係大麥。 (4)如第3項之液體麴之製法，其中大麥係上述碾製 比率爲90%以上者。 200538545 (5 )如第1或2項之液體麴之製法，其中榖類係米、 小麥、薔麥、稗、小米、稷、高梁或玉米。 (6 )如弟1項之液體麴之製法，其中只去除表面的榖 皮（粗殻）之榖類係糙米。 (7)如第1項之液體麴之製法，其中外皮覆蓋在表面 的未加工的丑類或芋類係大豆、小豆、或甘藷。 (8 )如第1〜7項中任一項之液體麴之製法，其中培養 於含上述培養原料的液體培養基之麴菌培養物中，至少同 _ 時生成、蓄積葡糖澱粉酶和耐酸性α -澱粉酶。 (9 ) 一種發酵飲食品之製法，其特徵係使用以上述1〜8 項中任一項方法製得的液體麹製造發酵飲食品。 (1 0 )如第9項之發酵飮食品之製法，其中發酵飲食品 的所有製造工程均在液態下進行。 (1 1 )如第9或1 0項之發酵飲食品之製法，其中發酵 飲食品的製造係於與外界保持遮蔽的液態狀態下進行。 (1 2 )如第9〜1 1項中任一項之發酵飲食品之製法，其 中發酵飲食品的製造係將原料置入上述液體麹中，製造一 _次酒醴。 (1 3 )如第9〜1 2項中任一項之發酵飲食品之製法，其 中發酵飲食品係燒酒。 (1 4 ) 一種發酵飲食品製造用的液體麴套組，其特徵係 至少含有以上述1〜8項中任一項之液體麴之製法製得的葡 糖澱粉酶活性和耐酸性α ·澱粉酶活性。 (1 5 )如第1〜7項中任一項之液體麴之製法，其中將麴 菌培養於含有上述培養原料的液體培養基以製造液體麴的 -10- 200538545 方法中，藉著抑制培養原料中澱粉內的糖質釋放至培養基 的速度，以調整液體麴的酵素活性。 〔發明效果〕 依據本發明，將麹菌培養於含有上述各種培養原料的液 體培養基，可製造液體麴，此液體麴可同時生成大量的葡 糖澱粉酶和耐酸性α -澱粉酶等釀造燒酒時所需的酵素 群。因液體培養較固體培養可更嚴密地監控培養條件，故 可經濟地且有效率地製造品質穩定之液體麴。 又，使用本發明製造的液體麴，具有和先前使用固體麴 ® 的燒酒酒醴相同程度的發酵性，如此製得的燒酒具有和使 用固體麴製得的燒酒相同程度的品質，可製得官能方面毫 不遜色之燒酒。 且，因本發明中使用的培養原料係未碾製、未加工、或 碾製程度至少爲外皮尙留在表面，故尙可提高原料利用率 或製品率。 又，使用本發明製得的液體麴製造燒酒時，不同於先前 使用固體麹製造燒酒的情形，因全工程可直接以液態進 •行，故可提供較先前更有效且穩定的燒酒製造系統。 【實施方式】 〔實施發明之最佳型態〕 以下，具體地說明本發明。 本發明的液體麴之製法係將麴菌培養於添加上述榖 類、豆類、芋類、特定的雜榖類等原料而調製成的液體培 養基，包括已提高葡糖激粉酶及耐酸性α -源粉酶的酵素活 性的液體麹之製造工程。亦即，因使用上述各種原料培養 -11- 200538545 麴菌，此原料中的澱粉的糖化時間長，可抑制糖被釋放至 培養基之速度，而提高液體麴的酵素活性。且葡糖澱粉酶 及耐酸性α -澱粉酶可同時地、平衡地生成並蓄積。 本發明中，使用爲原料的穀類例如大麥、米、小麥、蔷 麥、稗、小米、稷、高梁、玉米等。這些原料的型態，可 使用未碾製、或碾製比率爲至少有榖皮殘留在榖粒的表面 者等。例如榖類爲大麥時，未碾製的碾製比率1 00%者、或 以未碾製的碾製比率爲100%扣除大麥的榖皮比率（一般爲 7〜8 % )之比率，亦即碾製比率爲9 2〜9 3 %以上者。 ^ 上述的碾製比率，係指將榖類碾製後榖類殘留的比例， 例如碾製比率90%係指去除1 0°/。的榖類表層的榖皮等。 又，本發明中的糙麥，係指未碾製的麥至榖皮殘留在榖粒 表面的碾製程度間，亦即包含碾製比率9 0 %以上者。榖皮 係指覆蓋於榖粒表面的外側部分。 本發明中，使用爲原料的只去除表面的榖皮之榖類----糙米，係指只去除稻的粗殼的米。 又，本發明中使用爲原料的豆類和芋類，例如大豆、紅 •豆、甘藷等。這些原料只清洗外皮的污物，完全不施予剪 斷、粉碎處理等加工。 本發明中使用爲原料的莧紫係莧科莧屬植物的總稱，在 榖類中蛋白質含量高且胺基酸中的離胺酸的含量與大豆中 的含量相當。又，較碾製白米含更豐富的鈣、鐵、纖維質 等係高營養的縠類，原產國爲中南美洲諸國、印度、喜馬 拉雅、尼泊爾的特定區域。另一方面，奎藜籽係藜科的一 年生草本植物，主要栽培於祕魯南部或玻利維亞西部的安 -12- 200538545 地斯山脈等高地，富含礦物質、維生素、蛋白質、食物纖 維。 使用爲原料的莧紫和奎藜籽，可單獨使用或混合使用。 使用於液體培養基的調製時，不須進行細碎或粉碎等前處 理。 上述原料係和水混合調製成液體培養基。適當地選擇原 料的掺合比例，調製成液體培養基，使培養麹菌時可選擇 性地生成、蓄積葡糖澱粉酶及耐酸性α -澱粉酶。 例如以大麥作爲原料時，調製相對於水添加1〜20 %Biochem., 67, 2 1 94-2202 (2003) Non-Patent Literature 4. Sudo S. et al: J. Fernient. Bioeng., 76, 1 05- 1 1 0 (1 993) ^ Sudo S. et al: J. Ferment. Bioeng., 77, 48 3 -489 (1 994), Sudo Motojo, etc. · Japan Brewing Association Journal, 8 9,768 · 774 (1 994) [Summary] [Solve the problem of the invention] However, the patent In the method of reference 3, the culture of P. utilis with high activity of glucoamylase is produced by culturing P. utilis in a liquid medium prepared by adding a hardly decomposable saccharide, and it cannot be prepared by using raw materials such as pupae. Into ordinary liquid culture medium. In addition, the technology of cultivating oyster fungus in liquid medium to produce a high-activity glucoamylase yeast fungus culture is disclosed, but there is no mention of cultivating oyster fungus in a liquid medium to produce a highly active alcoholic fermentation. A key enzyme is the technology of acidic alpha-amylase-resistant liquid mash. It is generally considered that this acid-resistant α-amylase is an enzyme that cannot be produced in liquid culture, and so far, '尙 has not developed a highly active liquid which is resistant to acidic α-amylase'. The object of the present invention is to provide a method for producing liquid glutamate, which is characterized by the enzyme activity of glucoamylase and acid-resistant α-amylase, which are the key enzymes of alcoholic fermentation such as shochu, which are used in the production of fermented food and drink. Tall liquid maggots are cultivated using unmilled cereals, that is, maggots that cover the surface 200538545 or maggots (coarse shells) with only rinds removed from the surface, raw beans or taro, or specific cereals The liquid medium used as a raw material is not a special liquid medium cultured by adding a special sugar or the like, or using a raw material that has been roasted. [Methods to Solve the Problem] As a result of intensive research to solve the above-mentioned problems, the present inventors discovered that when manufacturing liquid tadpoles used in the production of fermented foods and drinks, the tadpoles were cultured as described above, and covered with any kind of tadpole Surface crickets or 去除 φ skins (coarse shells), peas or taro whose outer skin is covered on the surface, or hybrids such as osmanthus and / or quinoa (qUin〇a) A liquid culture medium used as a raw material for cultivation can be used to produce liquid mash with enhanced glucoamylase activity and acid resistance α_dianfenase activity. In addition, the liquid 麴 having enhanced enzyme activity is suitable for brewing shochu, and completed the present invention. That is, the present invention provides the following. (1) A method for producing liquid ravioli, which is characterized in that it is used for the production of liquid ravioli when fermenting foods and drinks. The fungus is cultivated on the surface of the ravioli that contains any kind of ravioli on the surface; ); Rinds; unprocessed beans or taro covering the surface; and citronella and / or quinoa seeds U ui η a) as a liquid culture medium. (2) The method of making liquid ravioli as described in item 1, wherein the ravioli whose skin is covered on the surface is unmilled or the milling ratio is such that at least the skin remains on the surface of the grain. (3) The method for producing liquid ravioli according to item 1 or 2, wherein the ravioli is barley. (4) The method for producing liquid ravioli according to item 3, wherein the above-mentioned milling ratio of barley is 90% or more. 200538545 (5) The method for making liquid ravioli according to item 1 or 2, wherein the ravioli is rice, wheat, rose wheat, coriander, millet, coriander, sorghum or corn. (6) The method for making liquid rice dumplings according to item 1, wherein only the rice dumplings (coarse shells) of the rice dumplings are removed from the surface. (7) The method for producing a liquid tincture according to item 1, wherein the surface is covered with raw ugly or taro soybeans, adzuki beans, or sweet potatoes. (8) The method for producing liquid ravioli according to any one of items 1 to 7, wherein the culture of radon bacterium in a liquid medium containing the above-mentioned culture raw material produces and accumulates glucoamylase and acid resistance at least at the same time Alpha-amylase. (9) A method for producing a fermented food or drink, which is characterized in that the fermented food or drink is manufactured by using liquid mash produced by any one of the methods of 1 to 8 above. (10) The method for producing fermented ravioli foods according to item 9, wherein all manufacturing processes of the fermented food and beverage are performed in a liquid state. (1 1) The method for producing a fermented food or drink as described in item 9 or 10, wherein the production of the fermented food or drink is carried out in a liquid state that is shielded from the outside. (1 2) The method for producing a fermented food or drink as described in any one of items 9 to 11, wherein the production of the fermented food or drink is made by placing raw materials in the above-mentioned liquid mash to produce one-time wine mash. (1 3) The method for producing a fermented food or drink according to any one of items 9 to 12, wherein the fermented food or drink is shochu. (1 4) A liquid tincture set for producing fermented food and drink, characterized in that it contains at least the glucoamylase activity and acid-resistant α · starch produced by the liquid tincture production method according to any one of 1 to 8 above. Enzyme activity. (1 5) The method for producing liquid radon according to any one of items 1 to 7, wherein the tadpole fungus is cultured in a liquid medium containing the above-mentioned culture raw material to produce liquid radon -10- 200538545, by inhibiting the culture raw material The rate at which the sugar in the starch is released to the medium to adjust the enzyme activity of the liquid mash. [Effects of the Invention] According to the present invention, liquid maggots can be produced by cultivating maggots in a liquid medium containing the above-mentioned various culture materials, and the liquid maggots can simultaneously produce a large amount of glucoamylase and acid-resistant α-amylase. Needed enzyme group. Because liquid culture can monitor the culture conditions more closely than solid culture, it is possible to economically and efficiently produce stable quality liquid puppets. In addition, the liquid 麴 produced by the present invention has the same degree of fermentability as the shochu 醴 using solid 麴 ®. The shochu produced in this way has the same quality as that of shochu made using solid 麴, and can be functionally prepared. No less shochu than that. In addition, since the culture raw materials used in the present invention are unmilled, unprocessed, or milled to a degree that at least the outer skin remains on the surface, the raw material utilization rate or product rate can be improved. In addition, when the shochu produced by using the liquid osmium produced by the present invention is different from the case where the shochu is produced by using the solid osmium, the entire process can be performed directly in a liquid state, so a more efficient and stable shochu production system can be provided. [Embodiment] [Best Mode for Carrying Out the Invention] Hereinafter, the present invention will be specifically described. The production method of the liquid ravioli of the present invention is to cultivate the fungi in a liquid medium prepared by adding the above-mentioned raw materials such as ravioli, beans, taro, and specific miscellaneous ravioli, including the improvement of glucokinase and acid resistance α- Manufacturing process of liquid peptone with enzyme activity derived from powder enzyme. That is, because the above-mentioned raw materials are used for cultivating -11-200538545 Pseudomonas spp., The starch in this raw material has a long saccharification time, which can inhibit the rate of sugar being released to the culture medium, and improve the enzyme activity of liquid pupae. In addition, glucoamylase and acid-resistant α-amylase can be produced and accumulated simultaneously and in balance. In the present invention, cereals such as barley, rice, wheat, rye, barley, millet, barley, sorghum, corn and the like are used as raw materials. The type of these raw materials may be unmilled or milled in such a manner that at least the skin of the crust remains on the surface of the crust. For example, when the barley is barley, the unmilled milling ratio is 100%, or the unmilled milling ratio is 100% minus the barley peel ratio (generally 7-8%), that is, The rolling ratio is 9 2 to 93% or more. ^ The above-mentioned milling ratio refers to the ratio of cricket residues after milling crickets. For example, the milling ratio of 90% means removing 10 ° /. The surface of the salamander skin and so on. In addition, the rye in the present invention refers to the degree of milling from the unmilled wheat to the surface of the coriander, which includes the milling ratio of 90% or more. Calyx skin refers to the outer part covering the surface of the calyx. In the present invention, brown rice, which is the raw material of which only the surface of the bark is removed, refers to rice from which only the coarse husk of rice is removed. The beans and taro used as raw materials in the present invention include, for example, soybeans, red beans, and sweet potatoes. These materials only clean the dirt on the outer skin, and are not subjected to processing such as cutting and pulverization. In the present invention, the genus Polygonum spp., Which is used as a raw material, is a general term for the genus Polygonum, which has a high protein content and a lysine content in amino acids that is equivalent to that in soybeans. In addition, compared with milled white rice, it contains richer calcium, iron, fiber, and other high-nutrition salamanders. The country of origin is a specific region of Central and South America, India, Himalaya, and Nepal. On the other hand, the annual herb of the quinoa seed family Chenopodiaceae is mainly cultivated in the highlands of Andes -12-200538545 in the south of Peru or western Bolivia, and is rich in minerals, vitamins, proteins, and food fiber. The osmanthus and quinoa seeds used as raw materials can be used alone or in combination. When used for the preparation of liquid culture media, no pretreatment such as fine crushing or pulverization is required. The above raw materials are mixed with water to prepare a liquid culture medium. The blending ratio of the raw materials is appropriately selected and prepared into a liquid medium, so that glucoamylase and acid-resistant α-amylase can be selectively produced and accumulated during the culture of P. utilis. For example, when using barley as a raw material, add 1 to 20% to water.

(w/vol )糙麥的液體培養基。又，使用未碾製的大麥作爲 糙麥時，調製成添加8〜10% ( w/vqI )的液體培養基較佳， 使用95%碾製的大麥作爲糙麥原料時，調製成添加1〜4% (w / v ο 1 )的液體培養基較理想。 其次，使用去粗殻的糙米作爲原料時，調製相對於水添 加1〜20% ( w/vol )的糙米的液體培養基，其中以添加5〜13 % ( w/vol )較理想，又以添加8〜10% ( w/vol )更佳。 使用豆類作爲原料時，調製相對於水添加1〜10% ( w/vol ) 的豆類的液體培養基，其中以添加8〜10% ( w/vol )較理想， 若使用紅豆時，調製添加1〜2% ( w/vol )的液體培養基。 又，使用芋類作爲原料時，調製相對於水添加1〜1 〇 % (w / v ο 1 )的芋類的液體培養基。 又，例如使用莧紫作爲原料時，調製相對於水添加 1.5〜15% ( w/vol)的液體培養基，其中以添加2〜10% ( w/vol) 較理想，又以添加2〜8% ( w/vol )更佳。另一方面，若使 用奎藜籽時，調製相對於水添加1 .5〜7% ( w/vol )的液體 -13- 200538545 培養基’其中以添加2〜6 % ( w / ν ο 1 )較理想，又以添加2〜4 % ( w / v ο 1 )更佳。 如此’最適當的掺合用量依使用原料的碾製度、使用的 麴菌株、原料的種類等而異，可任意地選擇。 將麴菌培養於添加適量的上述原料之液體培養基，平衡 地生成多量的葡糖澱粉酶及耐酸性α -澱粉酶，使用於燒酒 釀造可製得具有充分酵素活性的液體麴。原料的用量若超 過上限値，則培養液的黏性增加，無法充分供應麴菌進行 好氣培養所需的氧氣或空氣，培養物中的氧氣濃度降低， Β 培養過程不易繼續進行較不理想。另一方面，若此原料的 用量小於下限値，則無法提高酵素的產量。 亦可於培養前使原料中含有的澱粉預先糊化。使澱粉糊 化的方法無特別的限制，可採用蒸煮法、培炒法等常法。 後述的液體培養基的殺菌工程中，藉著高溫高壓滅菌等加 熱至高於澱粉的糊化溫度時，藉此處理亦可同時進行澱粉 的糊化。 在液體培養基中適當地添加上述原料之外的有機物、無 # 機鹽等營養源較理想。這些添加物可爲培養麹菌時一般使 用的物質，無特別的限制，有機物例如米糠、小麥麩、玉 米漿、大豆粕、脫脂大豆等，無機鹽例如銨鹽、硝酸鹽、 鉀鹽、酸性磷酸鹽、鈣鹽、鎂鹽等水溶性化合物，亦可同 時使用2種以上的有機物及/或無機鹽。其添加量係可促進 麴菌的增殖即可，無特別的限制，有機物的添加量爲0. 1〜5 % (w/vol)，無機鹽的添加量爲〇.1〜1% (w/vol)較理想。 如此製得的麴菌的液體培養基亦可依需求進行滅菌處理， -14- 200538545 其處理方法無特別的限制。例如可採用高溫高壓滅菌法， 於1 2 1°C進行1 5分鐘滅菌。 使滅菌後的液體培養基冷卻至培養溫度後，將麹菌接種 於液體培養基。本發明中使用的麴菌係具有糖質分解酵素 生成能之麹菌，又以具有葡糖澱粉酶生成能及耐酸性α -澱 粉酶生成能之麵菌較理想’例如以白麵菌（Aspergillus kawachii)等爲代表之白麴菌類、以泡盛麹菌（Aspergillus awamori)或黑麴菌（Aspergillus niger)等爲代表之泡盛麴 菌類、以黃麴菌（Aspergillus oryzae)、或醬油麹菌 (Aspergillus sojae)爲代表之黃麹菌類等。又，接種於培 養基的麴菌的型態不拘，可使用孢子或菌絲。 這些麴菌可由一種的菌株進行培養，或同種或二種以上 的異種菌株混合培養而成。可採用其孢子或由前培養收得 的菌絲等型態，採用菌絲的方法因達到對數增殖期所需時 間短故較理想。麴菌在液體培養基的接種量無特別的限 制，每lml液體培養基接種lxl 04〜lxl 〇6個孢子，若爲菌絲 則接種0. 1〜10%的前培養液。 麴菌的培養溫度，在不影響生育的範圍內無特別的限 制，惟以25〜45 °C較理想，又以30〜40°C更佳。若培養溫度 低則麹菌的增殖緩慢容易引起雜菌的污染。培養時間m 24〜72小時較理想。培養裝置係可進行液體培養者即可，惟 因麴菌必須進行好氣培養，必須於可將氧氣和空氣饋入培 養基中的好氣條件下進行。又，爲要在培養中使培養基中 的原料、氧氣、及麴菌可均勻地分布於裝置內，進行攪枠 較理想。攪拌條件和通氣量無特別的限制，可維持培養環 -15 - 200538545 境於好氣狀態即可，可依培養裝置、培養基的黏度等適當 地選擇。 藉由上述的培養法進行培養，可同時、平衡地生成葡糖 澱粉酶及耐酸性α -澱粉酶的酵素，形成具有可用於燒酒釀 造的酵素活性之液體麴。又，上述的培養法製得的液體麴， 除了培養物之外，亦可作爲使培養物離心分離收得的培養 液、其濃縮物或其乾燥物等。 由本發明的製法製得的液體麴等可用於酒類或發酵飲 食品的製造。例如製造清酒時在酒母或各種酒醴的釀造階 ® 段，製造燒酒時在酒醴的釀造階段，製造醬油時在釀造階 段，製造味噌時在釀造階段，製造味酣時在釀造階段，可 使用液體麴取代固體麹。 又，使用由上述液體麴或培養物收得的培養液或其濃縮 物等，製造酒類或發酵飲食品時，全部工程可以以液態進 行。全部工程以液態進行的酒類之製法，例如製造燒酒時， 使用玉米、麥、米、芋、稷等作爲原料，於8 0°C的高溫下， 使用耐熱性酵素劑使此原料溶解液化後，於其中再添加上 • 述液體麴及酵母，藉此使酒精發酵後的酒醴以常壓蒸留法 或減壓蒸留法等進行蒸留之製法。 〔實施例〕 以下，以實施例等更具體地說明本發明，惟本發明不受 限於這些實施例。 (實驗例1 )製造液體麴時糙麥的用量之檢討 調製如表1所示更改原料的糙麥比例之5種液體培養 基，將麴菌培養於各個液體培養基製造液體麴。 -16- 200538545 首先，在添加〇 . 2 % ( w / V ο 1 )硝酸鉀、0 · 3 % ( w / V ο 1 )磷 酸二氫鉀的水中加入糙麥，分別調製成含有1、2、4、8、 10% ( w/vol )糙麥的5種液體培養基。將l〇〇ml的調製成 的液體培養基放入容量5 00ml的附栓三角燒瓶中，在高溫 高壓滅菌釜中滅菌後，接種預先在液體培養基中進行前培 養的白麴菌（Aspergillus kawachii IF 0 4308)，其接種量相 對於液體培養基爲1% ( v/vol )。又，糙麥係使用國產2 條大麥的未碾製麥。 之後’於溫度3 7 °C、振盪速度1 〇 〇 r p m條件下培養4 8小 時。培養結束後，測定製得的各個培養物的葡糖澱粉酶、 耐酸性α -澱粉酶之生成量。表1及第1圖所示爲培養於不 同糙麥用量的液體培養基的培養物中，葡糖澱粉酶、耐酸 性α -澱粉酶之生成量。又，使用糖化力分別定量套組（龜 甲萬製）測定葡糖殿粉酶的酵素活性。又，耐酸性α -殿粉 酶的酵素活性之測定方法，係將（非專利文獻3 )的方法 稍加改良，培養物經過酸處理後使非耐酸性α -澱粉酶失活 後，使用α澱粉酶測定套組（龜甲萬製）進行測定。更具 體而言’在lml培養液中添加9ml的ι〇〇ηιΜ醋酸緩衝液 (ρΗ3 )，於37°C進行1小時酸處理後，使用以澱粉酶測 定套組（龜甲萬製）進行測定。 另方面’對照區係使用經碾製成碾製比率爲7 0 %的國 產2條大麥（以下稱爲圓麥）作爲原料，和試驗區相同做 法調製液體培養基，在相同條件下進行培養，培養結束後， 相同地測疋製得的各培養物的葡糖澱粉酶、耐酸性^ _澱粉 酶之生成里。表1及第2圖所示爲培養於不同圓麥用量的 -17- 200538545 令夜體培養基的培養物中，葡糖澱粉酶、耐酸性α -澱粉酶之 生成量。 (表1 ) ―原量和酵素生成量 試驗區 對照區 糙麥用量 GA (U/ml) ASAA (U/ml) 圓麥用量 GA (U/ml) AS AA (U/ml) 1% 72.4 3.1 1°/〇 58.8 1.2 2% 101.8 4.5 2% 102.6 3.4 4% 127.1 7.3 4% 66.2 4.2 8% 187.6 12.0 8% 17.5 11.2 10% 217.3 14.0 10% 10.3 10.7 16% 201.5 12.3 16% 10.0 9.7 20% 185.3 10.5 20% 8.8 8.9 * GA :葡糖澱粉酶 ASAA :耐酸性α _澱粉酶 如表1及第1圖所示，使用糙麥進行培養之培養物， 隨著糙麥用量的增加，葡糖澱粉酶和耐酸性α -澱粉酶的酵 素亦同時平衡地提高生成量，其生成量較使用圓麥的對照 區大幅地增加。特別是添加10% ( w/νοΐ )糙麥的液體培養 基，生成217.3 U/ml葡糖澱粉酶、14.0 U/ml耐酸性α -澱 粉酶，使用於燒酒製造時可同時具有充分的酵素活性。（僅 供參考：製造燒酒時所需的葡糖澱粉酶和耐酸性α -澱粉酶 的酵素活性値，葡糖澱粉酶爲l〇〇U/ml以上、耐酸性α -澱粉酶爲l〇U/ml以上） 另一方面，使用圓麥的對照區，如表1及第2圖所示， 添加2 % ( w / v ο 1 )圓麥的液體培養基的葡糖澱粉酶活性最 -18- 200538545 高，添加8% ( w/vol )圓麥的液體培養基的耐酸彳 酶活性最尚，二種酵素無法同時大量生成。 如此，將麴菌培養於添加1〜20% ( w/vol )糙 培養基可同時平衡地生成葡糖澱粉酶及耐酸伯 酶。特別是添加8〜10% ( w/voi )糙麥（未碾製） 使用於燒酒釀造亦無法製造同時具有充分的酵素 體麴。 將麴菌培養於使用糙麥的液體培養基後，可 地生成大量的葡糖澱粉酶及耐酸性α -澱粉酶，乃 皮覆蓋表面的糙麥作爲原料，藉著榖皮抑制原料 葡萄糖等糖的釋出，因於糖濃度較低的狀態下進 故較易生成葡糖澱粉酶及耐酸性α-澱粉酶等酵素 (實施例1 )使用糙麥的液體麴之製造 首先，在已添加0_2% ( w/vol )硝酸鉀、0.3% 磷酸二氫鉀的水中加入糙麥，調製成10% ( w/vo 培養基。其次，將l〇〇ml的液體培養基放入容量 附栓三角燒瓶，經過高溫高壓滅菌釜滅菌後，接 •液體培養基中進行前培養的白麴菌（Aspergillus IFO43 0 8 )，其接種量相對於液體培養基爲1% ( 又，糙麥係使用國產2條大麥的未碾製麥。 之後，於溫度37°C、振盪速度l〇〇rpm條件 小時。培養結束後，測定製得的各個培養物的葡糖 耐酸性α -澱粉酶之生成量，得知生成217.3 U/ml 酶、14.0 U/ml耐酸性α -澱粉酶，使用於燒酒製 時具有充分的酵素活性。 丨生α -澱粉 麥的液體 兰α -澱粉 時，即使 活性之液 同時平衡 因使用榖 中澱粉的 行培養， ； 〇 (w/vol) 1 )的液體 500ml 的 種預先在 kawachii v / v ο 1 ) 〇 下培養48 ί澱粉酶、 葡糖澱粉 造時可同 -19- 200538545 (實施例2 )以使用糙麥的液體麴製造麥燒酒 實施例1中，培養於添加(w/vol)糙麥調製成 的液體培養基而製得的液體麴（已增強葡糖澱粉酶和耐酸 性α -澱粉酶之培養物），使用此液體麴製造燒酒。 亦即，使用5 00ml的培養於添加10% ( w/vol )糙麥 調製成的液體培養基而製得的液體麴，如表2所示的掺合 釀造，進行麥合計量1328.6g的裝入釀造，發酵溫度維持 於25 °C，進行第一次釀造5日、第二次釀造2日、第三次 釀造13日共三段的裝入釀造。又，國產2條大麥經過70 %碾製並以水清洗後，浸漬60分鐘、瀝乾30分鐘後蒸煮 35分鐘之麥，使用此麥作爲續加入麥。又，第一次裝入釀 造中，因來自液體麹的帶入麥量50.0g無法充分進行發酵， 故裝入262.9g續加入麥使麥量和固體麴的麥量相同。又， 酵母係使用燒酒酵母（鹿兒島酵母），在30°C於YPD培養 基靜置培養4 8小時之培養物中植菌5 0 // 1。(w / vol) liquid medium for rye. In addition, when unmilled barley is used as brown wheat, it is preferable to prepare a liquid medium with 8 to 10% (w / vqI) added. When 95% milled barley is used as raw material for brown wheat, it is prepared to add 1 to 4 % (w / v ο 1) liquid medium is ideal. Secondly, when brown rice with husks removed is used as a raw material, a liquid medium in which 1 to 20% (w / vol) of brown rice is added to water is prepared. Among them, 5 to 13% (w / vol) is preferably added, and 8 to 10% (w / vol) is more preferable. When using beans as raw materials, prepare a liquid medium containing 1 to 10% (w / vol) of beans with respect to water. Among them, 8 to 10% (w / vol) is ideal. If red beans are used, add 1 to 2% (w / vol) liquid medium. When taro is used as a raw material, a liquid culture medium of taro added with 1 to 10% (w / v ο 1) with respect to water is prepared. For example, when using Violet Purple as a raw material, it is prepared to add 1.5 to 15% (w / vol) of liquid medium to water. Among them, 2 to 10% (w / vol) is preferably added, and 2 to 8% is added. (w / vol) is better. On the other hand, when using quinoa seeds, prepare 1.5-7% (w / vol) of liquid-13-200538545 medium with water added to it, in which 2 to 6% (w / ν ο 1) is added. Ideally, it is better to add 2 ~ 4% (w / v ο 1). In this way, the most appropriate blending amount varies depending on the milling system of the raw material used, the 麴 strain used, the type of the raw material, and the like, and can be arbitrarily selected. Pleurotus ostreatus is cultured in a liquid medium to which an appropriate amount of the above-mentioned raw materials are added, and a large amount of glucoamylase and acid-resistant α-amylase are produced in a balanced manner, and used for shochu brewing to produce liquid mash with sufficient enzyme activity. If the amount of the raw material exceeds the upper limit, the viscosity of the culture solution will increase, and the oxygen or air required for aerobic cultivation of the fungi can not be adequately supplied. The oxygen concentration in the culture will decrease, and it will be difficult to continue the cultivation process. On the other hand, if the amount of this raw material is less than the lower limit, the production of enzymes cannot be improved. The starch contained in the raw material may be gelatinized before the culture. The method for making the starch gelatinized is not particularly limited, and ordinary methods such as a cooking method and a frying method can be adopted. In the sterilization process of the liquid culture medium described later, when the temperature is higher than the gelatinization temperature of the starch by high-temperature autoclaving or the like, the gelatinization of the starch can be performed simultaneously by this treatment. It is desirable to appropriately add nutrient sources such as organic matter and organic salts other than the above-mentioned raw materials to the liquid culture medium. These additives can be generally used when cultivating tadpole bacteria, and there is no particular limitation. Organic materials such as rice bran, wheat bran, corn pulp, soybean meal, defatted soybeans, etc., inorganic salts such as ammonium, nitrate, potassium, acid phosphate Water-soluble compounds such as sodium, calcium, and magnesium salts may be used in combination of two or more organic and / or inorganic salts. The added amount is only required to promote the growth of P. spp., And is not particularly limited. The added amount of organic matter is 0.1 to 5% (w / vol), and the added amount of inorganic salt is 0.1 to 1% (w / vol) is ideal. The liquid culture medium of Pleurotus eryngii thus prepared can also be sterilized as required, and there is no particular limitation on the processing method. For example, high-temperature and high-pressure sterilization can be adopted, and sterilization is performed at 12 1 ° C for 15 minutes. After the sterilized liquid culture medium was cooled to the incubation temperature, the fungus was inoculated into the liquid culture medium. Pleurotus spp. Used in the present invention have glycolytic enzyme-producing ability. Pleurotus spp. Which has glucoamylase-producing ability and acid-resistant α-amylase-producing ability is more desirable. For example, Aspergillus kawachii The white fungus, such as Aspergillus awamori or Aspergillus niger, etc., are represented by Aspergillus awamori, Aspergillus niger, or Aspergillus oryzae, or Aspergillus sojae. Scutellaria baicalensis. In addition, the type of the fungus inoculated to the culture medium is not limited, and spores or hyphae can be used. These fungi can be cultured from one type of strain, or mixed cultures of the same type or two or more different types of strains. Types such as spores or hyphae obtained from pre-culture can be used. The method of hyphae is ideal because it takes a short time to reach the logarithmic proliferation period. 1 ~ 10% 的 前 cultivation liquid is inoculated in a liquid medium without specific limitation, and inoculated with lxl 04 ~ lxl 〇6 spores per lml of liquid medium. The culture temperature of Pleurotus spp. Is not particularly limited within a range that does not affect fertility, but 25 to 45 ° C is preferable, and 30 to 40 ° C is more preferable. If the culture temperature is low, the proliferation of tadpoles is slow and prone to contamination by miscellaneous bacteria. Incubation time m 24 ~ 72 hours is ideal. The culture device can be liquid culture, but the agaric bacteria must be aerobic cultured under aerobic conditions that can feed oxygen and air into the culture medium. In order to distribute the raw materials, oxygen, and bacillus in the culture medium uniformly in the device during the culture, it is preferable to stir. There are no particular restrictions on the agitation conditions and aeration volume, and the culture environment can be maintained -15-200538545 in an aerobic state, which can be appropriately selected according to the culture device and the viscosity of the culture medium. By culturing by the above-mentioned culturing method, glucoamylase and acid-resistant α-amylase-resistant enzymes can be produced simultaneously and in a balanced manner to form a liquid mash with enzyme activity that can be used for shochu brewing. The liquid mash produced by the above-mentioned culturing method can be used as a culture liquid obtained by centrifuging the culture, a concentrate thereof, or a dried product thereof, in addition to the culture. The liquid tincture and the like produced by the production method of the present invention can be used in the production of alcoholic beverages or fermented foods. For example, in the brewing stage of sake brewing or various sake brews when making sake, in the brewing stage when making shochu, in the brewing stage when making soy sauce, in the brewing stage when making miso, and in the brewing stage when making miso, you can use Liquid radon replaces solid radon. In addition, in the production of alcoholic beverages or fermented foods and drinks using the culture solution or the concentrate thereof obtained from the liquid mash or the culture, all processes can be performed in a liquid state. The whole process is to make liquor in liquid state. For example, when making shochu, corn, wheat, rice, taro, and coriander are used as raw materials, and the raw materials are dissolved and liquefied by using a heat-resistant enzyme at a high temperature of 80 ° C. The above-mentioned liquid mash and yeast are added to the method to make the mash with fermented alcohol after fermentation by normal pressure distillation or reduced pressure distillation. [Examples] Hereinafter, the present invention will be described more specifically with examples and the like, but the present invention is not limited to these examples. (Experimental Example 1) Review of the amount of brown wheat used in the production of liquid mash. Five types of liquid culture media were prepared by changing the ratio of rye as shown in Table 1. The mash was cultured in each liquid medium to produce liquid mash. -16- 200538545 First, add rye to water containing 0.2% (w / V ο 1) potassium nitrate and 0.3% (w / V ο 1) potassium dihydrogen phosphate to prepare 1, 2 , 4, 8, 10% (w / vol) of 5 types of liquid medium for rye. 100 ml of the prepared liquid culture medium was placed in a 500 ml capped Erlenmeyer flask, sterilized in an autoclave, and then inoculated with Aspergillus kawachii IF 0 previously cultured in the liquid culture medium. 4308), and its inoculation amount was 1% (v / vol) relative to the liquid medium. In addition, the rye line uses unmilled wheat made from two domestic barley. After that, the cells were cultured for 48 hours at a temperature of 37 ° C and an oscillation speed of 100 r p m. After the completion of the culture, the production amount of glucoamylase and acid-resistant α-amylase of each culture was measured. Tables 1 and 1 show the amounts of glucoamylase and acid-resistant α-amylase produced in cultures in liquid medium with different amounts of rye. In addition, the enzyme activity of glucosamine powder enzyme was measured using a saccharifying power quantitative set (Kikkoman). In addition, the method for measuring the enzyme activity of acid-resistant α-amylase is a slightly improved method (Non-Patent Document 3). After the culture is acid-treated, the non-acid-resistant α-amylase is inactivated, and α is used. The amylase measurement kit (Kikkoman) was used for measurement. More specifically, 9 ml of ιιιηιΜ acetic acid buffer solution (ρ 添加 3) was added to 1 ml of the culture medium, and after acid treatment at 37 ° C for 1 hour, the measurement was performed using an amylase measurement kit (made by Kikkoman). On the other hand, the control zone uses two domestic barleys with a milling ratio of 70% (hereinafter referred to as round barley) as raw materials. The liquid culture medium is prepared in the same way as in the test zone, and the culture is carried out under the same conditions. After the end, the production of glucoamylase and acid resistance ^ _amylase in each culture was similarly measured. Tables 1 and 2 show the amount of glucoamylase and acid-resistant α-amylase produced in cultures of -17-200538545 Lingye medium with different amounts of round wheat. (Table 1) ―The amount of raw wheat and the amount of enzyme production in the control area of the test area GA (U / ml) ASAA (U / ml) Round wheat GA (U / ml) AS AA (U / ml) 1% 72.4 3.1 1 ° / 〇58.8 1.2 2% 101.8 4.5 2% 102.6 3.4 4% 127.1 7.3 4% 66.2 4.2 8% 187.6 12.0 8% 17.5 11.2 10% 217.3 14.0 10% 10.3 10.7 16% 201.5 12.3 16% 10.0 9.7 20% 185.3 10.5 20% 8.8 8.9 * GA: Glucoamylase ASAA: Acid-resistant α_amylase As shown in Table 1 and Figure 1, cultures using rye for culturing. As the amount of rye increases, glucoamylase The enzyme and the acid-resistant α-amylase also increased the production amount in a balanced manner, and the production amount was significantly increased compared to the control area using round wheat. In particular, the liquid culture medium of 10% (w / νο 糙) rye was added to produce 217.3 U / ml glucoamylase and 14.0 U / ml acid-resistant α-amylase, which can have sufficient enzyme activity when used in the production of shochu. (For reference only: Enzymatic activity of glucoamylase and acid-resistant α-amylase required for the production of shochu. Glucoamylase is 100U / ml or more, and acid-resistant α-amylase is 10U. Above ml / ml) On the other hand, the control area using round wheat, as shown in Table 1 and Figure 2, shows that the glucoamylase activity of the liquid medium supplemented with 2% (w / v ο 1) round wheat is -18- 200538545 High, 8% (w / vol) liquid medium supplemented with round wheat has the most acid-resistant enzyme activity, and two enzymes cannot be produced in large quantities at the same time. In this way, culturing P. utilis in a medium supplemented with 1 to 20% (w / vol) coarse medium can simultaneously produce glucoamylase and primary acid-resistant enzymes in a balanced manner. In particular, the addition of 8 to 10% (w / voi) rye (unmilled) for shochu brewing does not produce sufficient enzymes. After cultivating P. utilis in a liquid medium using brown wheat, a large amount of glucoamylase and acid-resistant α-amylase can be produced. Rough wheat covered with skin is used as a raw material. It is released that it is easy to produce enzymes such as glucoamylase and acid-resistant α-amylase because it progresses at a low sugar concentration (Example 1) Production of liquid mash using rough wheat First, 0_2% has been added (w / vol) Potassium nitrate and 0.3% potassium dihydrogen phosphate were added to the wheat to prepare 10% (w / vo medium.) Next, 100 ml of liquid medium was placed in a volume-attached conical flask and subjected to high temperature After sterilization in an autoclave, the inoculated amount of Aspergillus IFO43 0 8 before being cultured in a liquid medium is 1% relative to the liquid medium. After that, the temperature was 37 ° C and the shaking speed was 100 rpm. After the cultivation was completed, the amount of glucose-resistant acidic α-amylase produced in each of the cultures was measured, and it was found that 217.3 U / ml was produced. Enzyme, 14.0 U / ml acid-resistant α-amylase, When used in shochu, it has sufficient enzyme activity. 丨 Liquid blue α-starch that produces α-starch wheat, even if the active liquid is balanced at the same time due to the cultivation of starch in 榖, 〇 (w / vol) 1) 500ml of liquid seed was cultured in advance under kawachii v / v ο 1) 48 ί amylase, glucostarch can be made the same as -19-200538545 (Example 2) to make barley shochu using liquid mash of rough wheat Example 1 The liquid mash (cultured with glucoamylase and acid alpha-amylase resistance) prepared by adding (w / vol) liquid medium prepared from rye to culture, was used to produce shochu. That is, 5,000 ml of liquid mash prepared by adding 10% (w / vol) rye prepared liquid culture medium was blended and brewed as shown in Table 2 to load 1328.6 g of wheat. Brewing, the fermentation temperature is maintained at 25 ° C, and the first brewing is carried out for 5 days, the second brewing for 2 days, and the third brewing for 13 days. In addition, after two domestic barleys were milled by 70% and washed with water, the wheat was dipped for 60 minutes, drained for 30 minutes, and then cooked for 35 minutes. This wheat was used as a continuous addition wheat. In the first brewing, the fermentation amount of 50.0 g of wheat brought in from the liquid mash did not sufficiently ferment, so 262.9 g of wheat was added and the amount of wheat was the same as that of the solid mash. In addition, the saccharomyces yeast (Kagoshima yeast) was used for the yeast system, and the plant was cultured at 30 ° C in a YPD culture medium for 4 to 8 hours, and the plants were planted with 50 0/1.

-20- 200538545 (表2 ) 試驗區（麥液體麹） 第一次 第二次 第三次 合計 麴麥（g) 50.0 一 — 50.0 續加入麥（g) 262.9 507.9 507.9 1278.6 液體麴（mL) 500.0 一 — 500.0 水（mL) 321.4 765.7 272.9 1360.0 90% 乳酸（mL) 1.4 一 一 1.4 又，對照裝入釀造（固體麴裝入釀造）係使用固體麴的 φ 麹麥，以表3所示的掺合釀造製造燒酒。固體麴之製法係 使用70%碾製麥，經洗麥後浸漬40分鐘，進行瀝乾30分 鐘，並蒸煮40分鐘後使冷卻至40 °C，對於每lkg碾製麥， 植菌 lg 的種麹（白麹菌 Aspergillus kawachii IFO4308)， 於40°C ·相對溼度95%的條件培養24小時，又於35°C · 相對溼度95%下培養6小時，再於30°C ·相對溼度90°/〇 下培養1 8小時。又，發酵條件等相同於上述本發明的裝入 釀造（液體麹裝入釀造）。 (表3 ) 對照區（麥固體麴） 第一次 第二次 第三次 合計 麴麥（g ) 312.9 312.9 續加入麥（β ) — 507.9 507.9 1015.7 水（mL ) 500.0 765.7 594.3 1860.0 第3圖所示係和對照組的固體麴裝入釀造之對照。從第 3圖可清楚得知，和使用固體麹的對照裝入釀造比較，在 使用液體麴的裝入釀造具有幾乎相同的發酵過程。又，無 -21- 200538545 論是使用液體麴或固體麴，其製得最後的酒_的酒精度數 皆爲1 7.8 % 。 其次，最後的酒醴經過減壓蒸留而得的原酒，加水使成 酒精度數爲25% ，由8位品評員以採點法（5點品評法、丄·· 佳〜5 :差）進行官能品評，其平均點如表4所示。 (表4 ) 燒酒原酒（酒精度數25% )之官能品評 _Wii_ 本發明裝入釀造（液體麴裝入釀浩、 3.0 對照裝入釀造（固體麴裝入釀浩） 3.0 其結果顯示，其酒質幾乎無差異性，無論使用液體麴或 固體麹皆可製得具相同酒質的燒酒。 從以上的結果得知，依據本發明將麴菌培養於添加丨〜2〇 % ( w/vol )糙麥的液體培養基，可平衡地同時生成葡糖澱 粉酶及耐酸性α -澱粉酶。特別是在添加8〜1〇% ( w/v〇1) 糙麥（未碾製）的液體培養基，可製得同時具有使用於燒 酒製造的充分的酵素活性之液體麴。因此，使用液體麴可 φ 製造和使用固體麹製得的燒酒相同酒質之燒酒。又，不須 以特別的培養裝置或特殊的培養工學方法嚴密地控制培 養，以簡便的液體培養基即可製造具有高活性葡糖澱粉酶 及耐酸性α ·澱粉酶之液體麴，且較固體培養容易地進行更 嚴密的製麴管理，可安定地製造高品質麴。又，藉由麴的 液體化不僅使酒醴的流動化的發酵管理簡易化，亦可達到 麹製造工程、燒酒製造工程的省力化、效率化。 (實施例3 )以使用蕎麥的液體麴製造米燒酒 -22- 200538545 1. 固體麴製造方法 使用9 0 %碾製米，洗米後浸漬15分鐘，瀝乾1 0分鐘， 並蒸煮30分鐘後使冷卻至40 °C，對於每lkg碾製米，植菌 lg 的種麴（白麴菌 Aspergillus kawachii IF0 4308)，於 40 °C •相對溼度9 5 %的條件培養2 4小時，又於3 5 °C ·相對 溼度9 5 %下培養6小時，再於3 0 °C ·相對溼度9 0 %下培養 1 8小時。 2. 液體麹之製法 (1 )前培養方法：8g的90%碾製米和l〇〇ml水放入 ® 5 0 0 m 1的附栓三角燒瓶，置於高溫高壓滅菌釜以1 2 1 °C進行 15分鐘滅囷。冷卻後’將白麹菌（Aspergillus kawachii IFO4308 )的種麹孢子植菌於前培養基中，使含量爲lxl〇6 個/m卜並於37°C，以lOOrpm震盪培養24小時。 (2)本培養法：將40g蔷麥和l.〇g硝酸鉀、i.5g磷酸 二氫鉀和5 00ml水放入2000ml的附栓三角燒瓶，置於高溫 尚壓滅囷盖以121C進丫了 15分鐘滅菌。將5ml前培養液植 菌於此本培養基，並於37 °C，以i〇〇r ριη震盪培養48小時， Φ 製造蔷麥液體麴。此時液體麴的酵素活性，GA活性爲 112.4U/ml ’ ASAA 活性爲 10.4U/nil。 3. 米燒酒之製法 (1)使用酵母：燒酒酵母（鹿兒島酵母） (2 )釀造掺合：釀造掺合如表5、表6所示。米係使用 90%碾製米，洗米後浸漬15分鐘，瀝乾1〇分鐘，並蒸煮 30分鐘後的米。試驗區分爲1)固體麴裝入釀造、2)薔麥 液體麴裝入釀造等2試驗區，二試驗區的總米量及水量相 -23- 200538545 等。酵母係將50 // 1於3(TC靜置培養48小時者植菌於YPD 培養基。 (3 )發酵條件：2 5 °C，固定 (4 )蒸留條件：減壓蒸留 (表5 ) 試驗區（喬麥液體麴） 第一次 第二次 第三次 合計 麴蕎麥（g) 40.0 一 一 40.0 續加入米（g ) 311.3 507.6 507.6 326.5 水（mL ) 594.0 765.4 265.6 1625.0 液體麴（mL) 500.0 一 0.0 500.0 90% 乳酸（mL) 1.4 一 — 1.4 (表6) 對照區（ 固體麴） 第一次 第二次 第三次 合計 麴米（g) 311.3 一 一 311.3 續加入米（g ) 一 507.6 507.6 1015.2 水（mL ) 594.0 765.4 565.6 2125.0 第4圖所示係和對照組的固體麴裝入釀造之對照。從第 4圖可清楚得知，和使用固體麴的對照裝入釀造比較，在 使用蕎麥液體麴的裝入釀造，具有幾乎相同的發酵過程。 又，固體麴裝入釀造區、蒿麥液體麴裝入釀造區的最後酒 醴的酒精度數分別爲1 9 .1 °/。、1 8 · 9 % ，幾乎相同程度。 其次，以減壓蒸留法使製得固體麹裝入釀造區和蔷麥液 體麹裝入釀造區的燒酒酒醴進行蒸留，由6位專業品評員 以5點品評法（佳3 - 5差）進行上述製得的燒酒原酒之官 -24- 200538545 能品評，結果可知固體麴裝入釀造區、喬麥液體麴裝入釀 造區無太大的差異，薔麥液體麴亦可製造具充分品質的燒 酒。又，如表7所示，薔麥液體麴區有「具喬麥風味」之 評語，可知使用喬麥液體麴可使米燒酒具有「蕎麥」般的 風味。 (表7 ) 官能品評結果 評點（平均點） 風評 固體麴 3.0 華麗 喬麥液體麴 2.8 舒暢，有喬麥風味 (實施例4 )以使用小米的液體麴製造米燒酒 1 ·固體麴製造方法 使用90%碾製米，洗米後浸漬15分鐘，瀝乾1〇分鐘， 並蒸煮30分鐘後使冷卻至4(TC，對於每lkg碾製米，植菌 lg 的種麴（白麹菌 Aspergillus kawachii IFO4308)，於 40 t ·相對溼度95%的條件培養24小時，又於351 ·相對 溼度95%下培養6小時，再於30°C ·相對溼度90%下培 養1 8小時。 2.液體麴之製法 (1 )前培養方法·· 8 g的9 0 %碾製米和1 〇 〇 m 1水放入 5 00ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121 °C進行 15分鐘滅菌。冷卻後，將白麴菌（Aspergillus kawachii IFO43 08 )的種麹孢子植菌於前培養基中，使含量爲lxl〇6 個/m卜並於37°C，以lOOrpm震盪培養24小時。 (2 )本培養法：將4 0 g小米和1 . 〇 g硝酸鉀、1 · 5 g磷酸 -25- 200538545 二氫鉀和5 0 0 m 1水放入2 0 0 0 m 1的附栓三角燒瓶，置於高溫 高壓滅菌釜以121°C進行15分鐘滅菌。將5ml前培養液植 菌於此本培養基，並於37°C，以lOOrpm震盪培養48小時， 製造小米液體麴。此時液體麹的酵素活性，GA活性爲 101.3U/ml，ASAA 活性爲 ll.OU/ml。 3 .米燒酒之製法 (1)使用酵母：燒酒酵母（鹿兒島酵母） (2 )釀造掺合：釀造掺合如表8、表9所示。米係使用 9 0 %碾製米，洗米後浸漬15分鐘，瀝乾1 〇分鐘，並蒸煮 30分鐘後的米。試驗區分爲1)固體麴裝入釀造、2)小米 液體麹裝入釀造等2試驗區’二試驗區的總米量及水量相 等。酵母係將5 0 // 1於3 0 °C靜置培養4 8小時者植菌於Y P D 培養基。 (3 )發酵條件：2 5 °C，固定 (4 )蒸留條件：減壓蒸留 (表8 ) 1 式驗區（小米液體麴） 第一次 第二次 第三次 合計 麴小米（g) 40.0 — 一 40.0 續加入米（g ) 311.3 507.6 507.6 1 326.5 液體麹（mL) 500.0 — — 500.0 水（mL ) 594.0 765.4 265.6 1 625.0 90% 乳酸（mL) 1.4 — — 1.4 -26· 200538545 (表9) -____ 對照區（固體麴） 第一次 第二次 第三次 a f+ 口口 1 麹米（g) 311.3 _ 311.3 續加入米（g) __ 507.6 507.6 1015.2 水（m L ) 594.0 765.4 565.6 2125.0 第5圖所示係和對照組的固體麴裝入釀造之對照。從第 5圖可、凊楚得知，和使用固體麴的對照裝入釀造比較，在 使用小米液體麴的裝入釀造，具有幾乎相同的發酵過程。 # 又’固體麴裝入釀造區、小米液體麴裝入釀造區的最後酒 醴的酒精度數均爲1 9.1 %相同。 其次’以減壓蒸留法使製得固體麹裝入釀造區和小米液 體麹裝入釀造區的燒酒酒醴進行蒸留，由6位專業品評員 以5點品評法（佳1-3-5差）進行上述製得的燒酒原酒之官 能品評，結果可知固體麴裝入釀造區和小米液體麹裝入釀 造區無太大的差異，小米液體麹亦可製造具充分品質的燒 酒。又，如表1 0所示，小米液體麴區有「舒暢的風味」之 φ 評語，顯示可製造和先前固體麴製法不同酒質的米燒酒原 酒0 -27- 200538545 (表 1 〇) 官能品評結果 評點（平均點） 風評 固體麴 3.0 華麗 小米液體麴 3.0 舒暢 (實施例5 )以使用稗的液體麴製造米燒酒 1 ·固體麴製造方法 使用90%碾製米，洗米後浸漬15分鐘，瀝乾10分鐘，並 蒸煮30分鐘後使冷卻至40 °C，對於每lkg碾製米，植菌 lg 的種麴（白麴菌 Aspergillus kawachii IFO4308)，於 40 _ °C ·相對溼度9 5 %的條件培養2 4小時，又於3 51 ·相對 溼度95%下培養6小時，再於30°C ·相對溼度90%下培 養1 8小時。 2 .液體麹之製法 (1 )前培養方法：8g的90%碾製米和100ml水放入5 00ml 的附栓三角燒瓶，置於高溫高壓滅菌釜以1 2厂C進行1 5分 鐘滅菌。冷卻後，將白麹菌（Aspergillus kawachii IFO4308) 的種麴孢子植菌於前培養基中，使含量爲1x1 〇6個/ml，並 於3 7 °c，以1 〇 〇 1· P m震盪培養2 4小時。 • (2)本培養法：將40g稗和l.Og硝酸鉀、1.5g磷酸二氫 鉀和5 00ml水放入2000ml的附栓三角燒瓶，置於高溫高壓 滅菌釜以1 2 1 °C進行1 5分鐘滅菌。將5 m 1前培養液植菌於 此本培養基，並於37°C，以lOOrpm震盪培養48小時’製 造稗液體麴。此時液體麴的酵素活性，GA活性爲 113.0U/ml，ASAA 活性爲 10.2U/ml。 3 .米燒酒之製法 (1)使用酵母：燒酒酵母（鹿兒島酵母） (2 )釀造掺合：釀造掺合如表1 1、表1 2所示。米係使用 -28- 200538545 9 0%碾製米，洗米後浸漬15分鐘，瀝乾l〇分鐘’並蒸煮 3 〇分鐘後的米。試驗區分爲1 )固體麴裝入釀造、2 )稗液 體麴裝入釀造等2試驗區，二試驗區的總米量及水量相 等。酵母係將5 0 // 1於3 0 °C靜置培養4 8小時者植囷Y P D 培養基。 (3 )發酵條件：2 5 °C，固定 (4 )蒸留條件：減壓蒸留 (表 1 1 ) 試驗區（稍 ί液體麴） 第一次 第二次 第三次 合計 麴稗（g) 40.0 一 — 40.0 續加入米（g) 311.3 507.6 507.6 1326.5 液體麴（mL) 500.0 — — 500.0 水（m L ) 594.0 765.4 265.6 1625.0 90% 乳酸（mL) 1.4 — — 1.4 (表 12) 對照區（固體麴） 第一次 第二次 第三次 合計 麹米（g) 311.3 — — 311.3 續加入米（g ) 一 507.6 507.6 1015.2 水（m L ) 594.0 765.4 765.6 2125.0 第6圖所示係和對照組的固體麹裝入釀造之對照。從第 6圖可清楚得知，和使用固體麴的對照裝入釀造比較，在 使用稗液體麴的裝入釀造，具有幾乎相同的發酵過程。又， 固體麴裝入釀造區、稗液體麴裝入釀造區的最後酒醴的酒 精度數分別爲1 9 .1 % 、1 8 · 8 %幾乎相同程度。 其次，以減壓蒸留法使製得固體麴裝入釀造區和稗液體 -29- 200538545 麹裝入釀造區的燒酒酒醴進行蒸留，由6位專業品評員以 5點品評法（佳1- 3 - 5差）進行上述製得的燒酒原酒之官能 品評，結果可知固體麴裝入釀造區、稗液體麴裝入釀造區 無太大的差異，稗液體麴亦可製造具充分品質的燒酒。又， 如表1 3所示，稗液體麴區有「舒暢的風味」之評語，顯示 可製造和先前固體麴製法不同酒質的米燒酒原酒。 (表 1 3 ) 官能品S平結果 評點（平均點） 風評 固體麴 3.0 華麗 稗液體麴 3.0 舒暢 (實施例6)以使用稷的液體麴製造米燒酒 1.固體麴製造方法 使用9 0 %碾製米，洗米後浸漬1 5分鐘，瀝乾1 〇分鐘， 並蒸煮30分鐘後使冷卻至40°C，對於每lkg碾製米，植菌 lg 的種麴（白麴菌 Aspergillus kawachii IF0 4308)，於 40 °C ·相對溼度95%的條件培養24小時，又於35 °C ·相對 φ 溼度95%下培養6小時，再於30°C ·相對溼度90%下培養 1 8小時。 2.液體麴之製法 (1 )前培養方法·· 8g的90%碾製米和100ml水放入 5 00ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121°C進行 15分鐘滅菌。冷卻後，將白麴菌（Aspergillus kawachii IFO43 0 8 )的種麴孢子植菌於前培養基中，使含量爲ΐχΐ〇δ 個/ m 1，並於3 7 °C，以1 0 0 r p m震盪培養2 4小時。 -30- 200538545 (2)本培養法：將40g稷和l.〇g硝酸鉀、1.5g磷酸二 氫鉀和5 00ml水放入2000ml的附栓三角燒瓶，置於高溫高 壓滅菌釜以1 2 1°C進行1 5分鐘滅菌。將5 ml前培養液植菌 於此本培養基，並於37°C，以lOOrpm震盪培養48小時， 製造稷液體麴。此時液體麴的酵素活性，GA活性爲 90.3U/ml，ASAA 活性爲 8.5U/m卜 3 .米燒酒之製法 (1)使用酵母：燒酒酵母（鹿兒島酵母） (2 )釀造掺合：釀造掺合如表14、表1 5所示。米係使 用9 0%碾製米，洗米後浸漬15分鐘，瀝乾10分鐘，並蒸 煮30分鐘後的米。試驗區分爲1)固體麴裝入釀造、2)稷 液體麴裝入釀造等2試驗區，二試驗區的總米量及水量相 等。酵母係將50 // 1於30°C靜置培養48小時者植菌於YPD 培養基。-20- 200538545 (Table 2) Test area (wheat liquid mash) Total rye (g) for the first, second, and third times 50.0 a — 50.0 Continue adding wheat (g) 262.9 507.9 507.9 1278.6 Liquid mash (mL) 500.0 a — 500.0 water (mL) 321.4 765.7 272.9 1360.0 90% lactic acid (mL) 1.4 one to one fourteen. Control brewing (solid 麴 loading brewing) is a φ rye barley using solid mash, brewed with the blend shown in Table 3. Making shochu. The method of making solid loquat is to use 70% milled wheat, immerse it for 40 minutes after washing it, drain it for 30 minutes, and cook it for 40 minutes to cool to 40 ° C. For each kg of milled wheat, plant lg species Centipede (Aspergillus kawachii IFO4308), cultured at 40 ° C · 95% relative humidity for 24 hours, and again at 35 ° C · 95% relative humidity for 6 hours, then at 30 ° C · 90% relative humidity Incubate for 18 hours. The fermentation conditions and the like are the same as those of the above-mentioned charging fermentation (liquid mash charging fermentation) of the present invention. (Table 3) Control area (wheat solid barley) Total barley (g) 312.9 312.9 Continue adding barley (β) — 507.9 507.9 1015.7 Water (mL) 500.0 765.7 594.3 1860.0 System shown in Figure 3 And the control group of solid tincture was filled into the brewing control. From Fig. 3, it is clear that compared with the control loading fermentation using solid mash, the fermentation using liquid mash has almost the same fermentation process. In addition, there is no -21-200538545 on the use of liquid rhenium or solid rhenium, and the alcohol content of the final wine produced is 1 7.8%. Secondly, the original wine obtained by decompression and distillation of the final sake wine was added with water to make the alcoholic content 25%, and 8 judges performed the function by using the picking method (5 points evaluation method, 丄 ·· Good ~ 5: poor). For evaluation, the average points are shown in Table 4. (Table 4) Functional evaluation of shochu original wine (alcohol content 25%) _Wii_ The present invention is brewed (liquid 麴 is loaded into brewing ho, 3.0 compared to the loading （(solid 麴 is loaded into brewing) 3.0) The results show that the wine There is almost no difference in quality, and shochu with the same wine quality can be produced regardless of the use of liquid 麹 or solid 麹. From the above results, it is known that 麴 bacteria are cultured in the addition of ～~ 20% (w / vol) according to the present invention. The liquid medium of rye can balance the simultaneous production of glucoamylase and acid-resistant α-amylase. Especially when adding 8 to 10% (w / v〇1) liquid medium of rye (unmilled), Liquid shochu with sufficient enzyme activity for shochu production can be produced. Therefore, using liquid shochu can produce shochu of the same quality as shochu produced using solid shochu. Also, no special cultivation device or Special cultivation engineering methods strictly control the cultivation, and can produce liquid 培养基 with high activity glucoamylase and acid-resistant α · amylase with simple liquid culture medium, and it is easier to carry out tighter 麴 management than solid culture Ke'an The production of high-quality ravioli is not only simplified, but also the fermentation management of fluidity of the ravioli is simplified by the liquidization of radon, and labor-saving and efficiency of the radon manufacturing process and shochu manufacturing process can be achieved. (Example 3) Production of rice shochu using buckwheat's liquid ravioli-22- 200538545 1. The method for producing solid ravioli uses 90% milled rice, wash the rice and soak it for 15 minutes, drain for 10 minutes, and cook for 30 minutes to cool to 40 ° C. For each 1 kg of milled rice, the seed pupae of plant lg (Aspergillus kawachii IF0 4308) are cultured at 40 ° C • 95% relative humidity for 2 4 hours, and at 3 5 ° C • relative humidity 9 Incubate at 5% for 6 hours, then incubate at 30 ° C and relative humidity for 90 hours at 18%. 2. Pre-cultivation method of liquid rice (1) Pre-cultivation method: 8g of 90% milled rice and 100ml Water was put into a conical flask with a plug of 5000 m 1 and placed in a high-temperature autoclave for 15 minutes at 1 2 ° C. After cooling, the spores of Aspergillus kawachii IFO4308 were planted. In the pre-culture medium, the content was 1 × 106 cells / m2 and cultured at 37 ° C. with shaking at 100 rpm for 24 hours. (2) This culture method: Put 40g of rosewood and 1.0g of potassium nitrate, i.5g of potassium dihydrogen phosphate and 500ml of water into a 2000ml Erlenmeyer flask with a stopper, put it in a high temperature and pressurized lid to 121C. It was sterilized for 15 minutes. 5 ml of the pre-culture broth was planted in this medium, and cultured at 37 ° C for 48 hours under shaking with i00r ριη. Φ produced rose barley liquid 此时. At this time, the enzyme activity of liquid 麴, GA activity was 112.4U / ml 'ASAA activity was 10.4U / nil. 3. Production method of rice shochu (1) Using yeast: Shochu yeast (Kagoshima yeast) (2) Brewing blending: The brewing blending is shown in Tables 5 and 6. Rice is 90% milled rice, washed for 15 minutes, drained for 10 minutes, and cooked for 30 minutes. The test is divided into 1) solid test) into brewing, 2) rose wheat liquid test into brewing, etc. 2 test areas, the total rice and water content of the second test area -23- 200538545 and so on. Yeast lined 50 // 1 at 3 (TC for 48 hours and cultured in YPD medium. (3) Fermentation conditions: 2 5 ° C, fixed (4) Steam retention conditions: vacuum distillation (Table 5) Test area (Qiaomai liquid tincture) The first, second and third times total buckwheat buckwheat (g) 40.0-40.0 Continue to add rice (g) 311.3 507.6 507.6 326.5 Water (mL) 594.0 765.4 265.6 1625.0 Liquid tincture (mL) 500.0-0.0 500.0 90% lactic acid (mL) 1.4 1 — 1.4 (Table 6) Control area (solid radon) Total rice (g) for the first, second and third times 311.3-311.3 Continue to add rice (g)-507.6 507.6 1015.2 Water (mL) ) 594.0 765.4 565.6 2125.0 The solid mash of the system and control group shown in Figure 4 is loaded into the brewing control. As can be clearly seen from Figure 4, compared with the control using solid mash into the brewing, The brewing process has almost the same fermentation process. In addition, the alcohol content of the final wine tartare in the solid wine tartare in the brewing area and the wormwood liquid tartare in the brewing area are 19.1 ° /., 18 · 9%, Almost the same degree. The solid 麹 is loaded into the brewing area and the rose barley liquid 麹 is loaded into the shochu wine 醴 in the brewing area for steaming and retention. Six professional tasters use the 5-point evaluation method (good 3-5 difference) to perform the above-mentioned official shochu wine production- 24- 200538545 It can be evaluated. The results show that there is not much difference between the solid 麴 loading into the brewing zone and the Qiaomai liquid 麴 loading into the brewing zone, and the rose wheat liquid 麴 can also produce shochu with sufficient quality. Also, as shown in Table 7, There is a comment on "Qiaomai flavor" in the rose wheat liquid area. It can be seen that the use of the Qiaomai liquid type can make rice shochu have a "buckwheat" flavor. (Table 7) Functional evaluation result evaluation points (average point) Wind evaluation solid status 3.0 gorgeous Qiaomai liquid 麴 2.8 is comfortable and has Qiaomai flavor (Example 4) to make rice shochu using millet's liquid 麴 1. The solid 麴 manufacturing method uses 90% milled rice, rinsed for 15 minutes, and drained for 10 minutes. After 30 minutes of cooking, it was cooled to 4 ° C (TC, for 1 kg of milled rice, the species of Aspergillus kawachii IFO4308) was cultured at 40 t · 95% relative humidity for 24 hours, and then at 351 · Relative humidity 95% After 6 hours of cultivation, incubate at 30 ° C for 18 hours at 90% relative humidity. 2. Pre-cultivation method of liquid mash (1) · 8 g of 90% milled rice and 100m 1 water Put in a 500 ml conical Erlenmeyer flask with a stopper and place in an autoclave at 121 ° C for 15 minutes. After cooling, the Aspergillus kawachii IFO43 08 seed spores were planted in the pre-culture medium at a content of 1 × 106 cells / m and cultured at 37 ° C. with shaking at 100 rpm for 24 hours. (2) This culture method: Put 40 g of millet and 1.0 g of potassium nitrate, 1.5 g of phosphoric acid-25- 200538545 potassium dihydrogen and 5000 m 1 of water into the plug of 2000 m 1 The conical flask was placed in a high-temperature autoclave and sterilized at 121 ° C for 15 minutes. 5 ml of the former culture liquid was planted in this medium, and cultured at 37 ° C. with shaking at 100 rpm for 48 hours to produce millet liquid mash. At this time, the enzyme activity of the liquid puppet, GA activity was 101.3 U / ml, and ASAA activity was ll.OU / ml. 3. Production method of rice shochu (1) Using yeast: Shochu yeast (Kagoshima yeast) (2) Brewing blending: Brewing blending is shown in Tables 8 and 9. Rice is 90% milled rice, washed for 15 minutes, drained for 10 minutes, and cooked for 30 minutes. The tests were divided into 1) solid 麴 loading and brewing, 2) millet liquid 麹 loading and brewing, etc. 2 test zones', the total rice and water contents were equal. Yeast lines were cultured for 5 8 // 1 at 30 ° C for 4 to 8 hours, and then planted in Y P D medium. (3) Fermentation conditions: 2 5 ° C, fixed (4) Distillation conditions: Decompression distillation (Table 8) Type 1 inspection area (Xiaomi liquid 麴) Total 第二 Xiaomi (g) 40.0 — 1 40.0 Continue to add rice (g) 311.3 507.6 507.6 1 326.5 Liquid tincture (mL) 500.0 — — 500.0 Water (mL) 594.0 765.4 265.6 1 625.0 90% lactic acid (mL) 1.4 — — 1.4 -26 · 200538545 (Table 9) -____ Control area (solid radon) First time Second time Third time a f + Mouth 1 Rice (g) 311.3 _ 311.3 Continue to add rice (g) __ 507.6 507.6 1015.2 Water (m L) 594.0 765.4 565.6 2125.0 Figure 5 shows The solid pupae of the line and the control group were loaded into the brewing control. From Fig. 5, we can see that compared with the control loading brewing using solid tincture, the loading fermentation using millet liquid tincture has almost the same fermentation process. # 又 ’The solid alcohol is loaded into the brewing area, and the millet liquid liquid is loaded into the brewing area. The alcohol content of the final wine is 19.1%. Secondly, the solid 麹 produced by the reduced pressure steaming method was loaded into the brewing area and the millet liquid was loaded into the shochu wine liquor in the brewing area for steaming and retention. Six professional judges evaluated the method by 5 points (good 1-3-5 difference) ) According to the functional evaluation of the shochu raw liquor prepared above, it can be seen that there is not much difference between the solid 麴 loading in the brewing zone and the Xiaomi liquid 麹 loading in the brewing zone, and the Xiaomi liquid 麹 can also produce shochu with sufficient quality. In addition, as shown in Table 10, Xiaomi's liquid food zone has a φ comment of "smooth flavor", showing that rice shochu original sake that can be produced with a different quality from the previous solid production method is 0 -27- 200538545 (Table 1 〇) Functional evaluation Result evaluation point (average point) Wind evaluation solid 麴 3.0 gorgeous millet liquid 麴 3.0 comfortable (Example 5) to make rice shochu with liquid 麴 using 稗 1 · solid 麴 manufacturing method uses 90% milled rice, immerse for 15 minutes after washing the rice, drain Dry for 10 minutes and cook for 30 minutes to cool to 40 ° C. For each 1 kg of milled rice, plant lg seed worm (Aspergillus kawachii IFO4308), at 40 _ ° C · 95% relative humidity Conditioned cultivation was performed for 24 hours, and then incubated at 3 51 · relative humidity for 95% for 6 hours, and then at 30 ° C · relative humidity for 90% for 18 hours. 2. Preparation method of liquid tincture (1) Pre-cultivation method: 8g of 90% milled rice and 100ml of water are put into a 500ml conical Erlenmeyer flask, placed in a high-temperature autoclave and sterilized at 12C for 15 minutes. After cooling, the Aspergillus kawachii IFO4308 spore spores were planted in the pre-culture medium to a content of 1 × 10 6 / ml, and cultured at 37 ° C. with a vibration of 1,000 · P m 2 4 hours. • (2) This culture method: Put 40 g of tadpoles and 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate and 500 ml of water into a 2000 ml conical Erlenmeyer flask, and place in a high-temperature autoclave at 1 2 1 ° C Sterilize for 15 minutes. 5 ml of the pre-culture broth was planted in this medium, and cultured at 37 ° C with shaking at 100 rpm for 48 hours to produce a mash liquid. At this time, the enzyme activity of the liquid puppet, GA activity was 113.0 U / ml, and ASAA activity was 10.2 U / ml. 3. Production method of rice shochu (1) Using yeast: Shochu yeast (Kagoshima yeast) (2) Brewing blending: Brewing blending is shown in Table 1 and Table 12. For rice, use -28- 200538545 9 0% milled rice, wash the rice and soak for 15 minutes, drain for 10 minutes' and cook for 30 minutes. The test is divided into 2) test areas, 1) solid 麴 loading and brewing, 2) liquid 麴 loading and brewing, etc. The total rice and water content in the second test zones are equal. Yeast lines were cultured at 50 0 // 1 at 30 ° C for 4 to 8 hours, and planted with Y P D medium. (3) Fermentation conditions: 2 5 ° C, fixed (4) Distillation conditions: Decompression distillation (Table 1 1) Test area (slightly liquid) First, second, and third total 麴 稗 (g) 40.0 40.0 Continue to add rice (g) 311.3 507.6 507.6 1326.5 Liquid tritium (mL) 500.0 — — 500.0 Water (m L) 594.0 765.4 265.6 1625.0 90% lactic acid (mL) 1.4 — 1.4 (Table 12) Control area (solid tritium) The first time, the second time and the third time, total rice (g) 311.3 — — 311.3 Continue to add rice (g)-507.6 507.6 1015.2 Water (m L) 594.0 765.4 765.6 2125.0 The solid radon shown in Figure 6 and the control group is loaded into the brewing The contrast. It is clear from FIG. 6 that compared with the control loading fermentation using solid mash, the fermentation using 稗 liquid 麴 loading has almost the same fermentation process. In addition, the solid wines were loaded into the brewing zone, and the liquid wines were loaded into the final wine of the brewing zone. The wine accuracy numbers were almost 19.1% and 18 · 8%, respectively. Secondly, the prepared solid 麴 was loaded into the brewing area and 稗 liquid -29- 200538545 by the reduced pressure distillation method. The shochu wine 麹 loaded into the brewing area was steamed and retained by 6 professional judges with a 5-point evaluation method (佳 1- 3-5 difference) The functional evaluation of the shochu original liquor prepared above shows that there is not much difference between solid 麴 loading in the brewing zone and 稗 liquid 麴 loading in the brewing zone. 稗 Liquid 麴 can also produce shochu with sufficient quality. In addition, as shown in Table 13, the "liquid" region has a "smooth flavor" comment, indicating that it is possible to produce rice shochu liquor that is different from the previous solid method. (Table 1 3) Functional product S-level result evaluation points (average point) Wind evaluation solid 麴 3.0 gorgeous 稗 liquid 麴 3.0 comfortable (Example 6) rice shochu made with liquid 麴 using 稷 1. solid 麴 manufacturing method using 90% mill For rice making, immerse for 15 minutes after washing, drain for 10 minutes, and cook for 30 minutes to cool to 40 ° C. For each kg of milled rice, plant lg seed worm (Aspergillus kawachii IF0 4308) Incubate at 40 ° C · 95% relative humidity for 24 hours, then at 35 ° C · relative φ 95% humidity for 6 hours, then at 30 ° C · 90% relative humidity for 18 hours. 2. Production method of liquid mash (1) Pre-cultivation method: 8g of 90% milled rice and 100ml of water are put into a 500ml conical flask with a plug, and placed in a high-temperature autoclave for 121 minutes at 15 ° C for sterilization. After cooling, the spores of Aspergillus kawachii IFO43 0 8 were planted in the pre-culture medium to a content of ΐχΐδδ / m 1 and cultured at 37 ° C with shaking at 100 rpm. 24 hours. -30- 200538545 (2) The culture method: Put 40 g of ytterbium and 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 ml of water into a 2000 ml conical Erlenmeyer flask, and place in a high-temperature autoclave to 1 2 Sterilize at 1 ° C for 15 minutes. 5 ml of the pre-culture broth was planted in this medium, and cultured at 37 ° C. with shaking at 100 rpm for 48 hours to produce 稷 liquid 麴. At this time, the enzyme activity of liquid mash, GA activity was 90.3U / ml, ASAA activity was 8.5U / m3. The method for making rice shochu (1) using yeast: shochu yeast (Kagoshima yeast) (2) brewing blending: The brewing blends are shown in Tables 14 and 15. Rice is 90% milled rice, washed for 15 minutes, drained for 10 minutes, and cooked for 30 minutes. The tests are divided into 2) test areas, 1) solid 麴 loading and brewing, 2) 麴 liquid 等 loading and brewing, etc. The total rice and water content in the two test zones are equal. Yeast strains were cultured at 50 // 1 at 30 ° C for 48 hours and planted in YPD medium.

(3)發酵條件：25°C，固定 (4 )蒸留條件：減壓蒸留 (表 14 ) 試驗區（稷液體麴） 第一次 第二次 第三次 合計 麴稷（g) 40.0 _ _ 40.0 續加入米（g) 311.3 507.6 507.6 1326.5 液體麴（mL) 500.0 — ___ 500.0 水（m L ) 594.0 765.4 265.6 1625.0 90% 乳酸（mL) 1.4 — 一 1.4 200538545 (表 15 ) 對照區（固體麴） 第一次 第二次 第三次 合計 麹米（g) 311.3 一 — 311.3 續加入米（g) — 507.6 507.6 1015.2 水（m L ) 594.0 765.4 765.6 2125.0 第7圖所示係和對照組的固體麴裝入釀造之對照。從第 7圖可清楚得知，和使用固體麴的對照裝入釀造比較，在 使用稷液體麴的裝入釀造，具有幾乎相同的發酵過程。又， φ 固體麴裝入釀造區和稷液體麴裝入釀造區的最後酒醴的酒 精度數分別爲1 9 · 1 % 、1 8 · 8 %幾乎相同程度。 其次，以減壓蒸留法使製得固體麴裝入釀造區和稷液體 麹裝入釀造區的燒酒酒醴進行蒸留，由6位專業品評員以 5點品評法（佳1-3-5差）進行上述製得的燒酒原酒之官能 品評，結果可知固體麹裝入釀造區、稷液體麴裝入釀造區 無太大的差異，稷液體麴亦可製造具充分品質的燒酒。又’ 如表1 6所示，稷液體麴區有「具圓滑、甘味的味道」之評 語，顯示可製造和先前固體麴製法不同酒質的米燒酒原酒。 -32- 200538545 (表 16 ) 官能品g平結果 評點（平均點） 風評 固體麴 3.0 華麗 稷液體麴 2.9 甘味、圓滑 (實施例7 )以使用高梁的液體麴製造米燒酒 1 .固體麹製造方法 使用90%碾製米，洗米後浸漬15分鐘，瀝乾10分鐘， 並蒸煮30分鐘後使冷卻至40 °C，對於每lkg碾製米，植菌 _ lg 的種麴（白麴菌 Aspergillus kawachii IFO4308)，於 40 °C ·相對溼度95%的條件培養24小時，又於35 °C ·相對 溼度95%下培養6小時，再於30°C ·相對溼度90%下培 養1 8小時。 2.液體麹之製法 (1 )前培養方法：8g的90%碾製米和1 〇〇ml水放入 5 00ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121 °C進行 15分鐘滅菌。冷卻後，將白麴菌（Aspergillus kawachii φ IFO43 0 8 )的種麹孢子植菌於前培養基中，使含量爲1χ106 個/ml，並於37°C，以lOOrpm震盪培養24小時。 (2 )本培養法：將40g高梁和l.Og硝酸鉀、1.5g磷酸 二氫鉀和5 0 0 m 1水放入2 0 0 0 m 1的附栓三角燒瓶，置於高溫 高壓滅菌釜以1 2 1 °C進行1 5分鐘滅菌。將5 m 1前培養液植 菌於此本培養基，並於37°C，以l〇〇rpm震盪培養48小時， 製造高梁液體麴。此時液體麴的酵素活性，GA活性爲 111.2U/ml，ASAA 活性爲 l〇.5U/ml。 -33- 200538545 3 .米燒酒之製法 (1 )使用酵母··燒酒酵母（鹿兒島酵母） (2 )釀造掺合：釀造掺合如表丨7、表1 8所示。米係使 用9 0 %碾製米，洗米後浸漬1 5分鐘，瀝乾1 〇分鐘，並蒸 煮30分鐘後的米。試驗區分爲1)固體麴裝入釀造、2) 高粱液體麴裝入釀造等2試驗區，二試驗區的總米量及水 量相等。酵母係將50 // 1於30°C靜置培養48小時者植菌於 YPD培養基。 (3 )發酵條件：2 5 °C (4 )蒸留條件：減壓蒸留 (表 17 ) 試驗區（高粱液體麴） 第一次 第二次 第三次 合計 麹局粱（g ) 40.0 一 一 40.0 續加入米（g) 311.3 507.6 507.6 1326.5 液體麵（m L ) 500.0 _ 一 500.0 水（mL ) 594.0 765.4 265.6 1625.0 90% 乳酸（mL) 1.4 — — 1.4 (表 1 8 ) 對照區（固體麴） 第一次 第二次 第三次 合計 麹米（g) 311.3 — 一 311.3 續加入米（g 1 _ 507.6 507.6 1015.2 水（mL) 594.0 765.4 765.6 2125.0 第8圖所示係和對照組的固體麴裝入釀造之對照。從第 圖可清楚得知，和使用固體麴的對照裝入釀造比較，在 -34- 200538545 使用高粱液體麴的裝入釀造，具有幾乎相同的發酵過程。 又’固體麴裝入釀造區、高梁液體麴裝入釀造區的最後酒 醴的酒精度數均爲1 9 . 1 %相同程度。 其次，以減壓蒸留法使製得固體麴裝入釀造區和高梁液 體麴裝入釀造區的燒酒酒醴進行蒸留，由6位專業品評員 以5點品評法（佳1-3-5差）進行上述製得的燒酒原酒之官 能品評，結果可知固體麴裝入釀造區、高粱液體麴裝入釀 造區無太大的差異，高梁液體麴亦可製造具充分品質的燒 酒。又，如表1 9所示，高梁液體麴區有「具穀物的甘味」 ^ 之評語，顯示可製造和先前固體麴製法有明顯差異化的燒 酒原酒。 (表 1 9 ) 官能品評結果 評點（平均點） 風評 固體麹 3.0 華麗 高梁液體麴 3.0 穀物的甘味 (實施例8)使用玉米的液體麴之製造 (1 )前培養方法：8 g的9 0 %碾製米和10 0 m 1水放入 5 00ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121°C進行 15分鐘滅菌。冷卻後，將白麴菌（Aspergillus kawachii IFO4308)的種麴孢子植菌於前培養基中，使含量爲1χ1〇6 個/ml，並於37°C，以lOOrpm震盪培養24小時。 (2 )本培養法：將1〜8g玉米和0.2g硝酸鉀、〇.3g磷酸 二氫鉀和100ml水放入500ml的附栓三角燒瓶’置於高溫 高壓滅菌釜以1 2 1°C進行1 5分鐘滅菌。將1 ml前培養液植 -35- 200538545 菌於此本培養基’並於37°C，以l〇〇rpm震盪培養48小時。 利用實驗例1的方法測定培養後的培養上清液中的酵素生 成量，亦即葡糖澱粉酶（G A )活性及耐酸性α -澱粉酶 (A S A A )活性。結果如表2 0所示。 從表2 0清楚可知’在玉米用量4 %以上驗區，清楚得知 燒酒釀造所需的酵素活性的目標値葡糖澱粉酶清楚爲 100U/ml。另一方面，耐酸性α -澱粉酶的目標値爲i〇u/m卜 隨者增加無法達到此目標値區的玉米用量，此酵素的生成 量有增加的趨勢。 如上述般，本試驗的AS AA活性的目標値不清楚者，因 具有同時生成GA酵素、及AS A A酵素之能力，極可能藉著 培養條件的最適化使酵素生成性增大至目標値水準。又， 在使用8 %玉米液體麴的燒酒裝入釀造中，若較一般掺合 增加麴比率，可充分地製造燒酒。 (表 20 ) 玉米用量 酵素活性（U/ml) GA ASAA 1°/〇 61.9 1.4 2% 66.4 1.7 4% 135.7 3.8 8% 114.2 4.2 (實施例9)使用糙麥的液體麴之製造 (1 )前培養方法：8g的65%碾製麥和l〇〇ml水放入 5 00ml的附栓三角燒瓶，置於高溫高壓滅菌釜以12 1°C進行 1 5分鐘滅菌。冷卻後，將泡盛麴菌（A s p e r g i 11 u s a w a m 〇 r i IF0 4388 )的種麴孢子植菌於前培養基中，使含量爲lxl〇6 -36- 200538545 個/ml，並於37°C，以lOOrpm震盪培養24小時。 (2 )本培養法：將1〜8g糙麥（95%碾製麥）和〇.2g硝 酸鉀、0.3g磷酸二氫鉀和100ml水放入500ml的附栓三角 燒瓶’置於高溫高壓滅菌釜以1 2 1°C進行1 5分鐘滅菌。將 lml前培養液植菌於此本培養基，並於37 °C，以lOOrpm震 盪培養4 8小時。利用實驗例1的方法測定培養後的培養上 清液中的酵素生成量，亦即葡糖澱粉酶（GA )活性及耐酸 性α -澱粉酶（A S A A )活性。結果如表2 1所示。 從表2 1清楚可知，在糙麥用量爲4%的試驗區，得知燒 酒釀造所需的酵素活性的目標値葡糖澱粉酶清楚爲 1 0 0U/ml及耐酸性α -澱粉酶清楚爲1 OU/ml。由此可知，即 使使用黑麹菌仍具有和白麴菌相同的酵素的高生成效果。 (表 21 ) 糙麥用量 酵素活性（U/ml) GA ASAA 1% 27.9 2.6 2% 41.5 2.0 4% 136.5 10.0 8°/〇 7.1 0.1(3) Fermentation conditions: 25 ° C, fixed (4) Distillation conditions: Decompression distillation (Table 14) Test area (稷 Liquid 麴) First, second, and third total g (g) 40.0 _ _ 40.0 Continue adding Rice (g) 311.3 507.6 507.6 1326.5 Liquid radon (mL) 500.0 — ___ 500.0 Water (m L) 594.0 765.4 265.6 1625.0 90% Lactic acid (mL) 1.4 — 1.4 1.438538545 (Table 15) Control area (solid radon) First The second and third times total indica rice (g) 311.3-311.3 Continue to add rice (g) — 507.6 507.6 1015.2 Water (m L) 594.0 765.4 765.6 2125.0 The solid osmium shown in Figure 7 and the control group is loaded into the brewing control . It is clear from FIG. 7 that, compared with the control loading brewing using solid mash, the brewing using 稷 liquid 麴 has almost the same fermentation process. In addition, the precision of φ solid 麴 loaded into the brewing zone and 稷 liquid 麴 loaded into the final wine of the brewing zone were 19 · 1% and 18 · 8%, respectively, to almost the same degree. Secondly, the solid 麴 produced in the brewing area and the 稷 liquid 麹 in the brewing area were steamed and retained by the reduced pressure steam retention method. Six professional judges evaluated the method by 5 points (good 1-3-5 difference) ) According to the functional evaluation of the shochu raw liquor prepared above, it can be seen that there is not much difference between the solid 麹 loading into the brewing zone and the 稷 liquid 麴 loading into the brewing zone, and the 稷 liquid 麴 can also produce shochu with sufficient quality. Also, as shown in Table 16, the 稷 liquid 麴 area has the comment of “smooth, sweet taste”, indicating that it is possible to produce rice shochu liquor with a different quality from the previous solid 麴 method. -32- 200538545 (Table 16) Functional product g-level result evaluation point (average point) Wind rating solid 麴 3.0 gorgeous 稷 liquid 麴 2.9 sweet and smooth (Example 7) to use rice sorghum liquid 麴 to produce rice shochu 1. solid 麹 manufacturing method Use 90% milled rice, soak the rice for 15 minutes, drain it for 10 minutes, and cook for 30 minutes to cool to 40 ° C. For each kg of milled rice, plant _ lg of seed worm (Aspergillus kawachii) IFO4308), incubate at 40 ° C · 95% relative humidity for 24 hours, then at 35 ° C · 95% relative humidity for 6 hours, and then at 30 ° C · 90% relative humidity for 18 hours. 2. Preparing method of liquid mash (1) Pre-cultivation method: 8g of 90% milled rice and 1000ml of water are put into a 500ml Erlenmeyer flask with a plug and placed in a high-temperature autoclave for 121 minutes at 15 ° C for sterilization . After cooling, Aspergillus kawachii φ IFO43 0 8 spores were planted in the pre-culture medium to a content of 1 × 106 cells / ml, and cultured at 37 ° C. with shaking at 100 rpm for 24 hours. (2) The culture method: Put 40 g of sorghum and 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate, and 500 m 1 of water into a 2000 m 1 attached Erlenmeyer flask, and place in a high-temperature autoclave Sterilize at 1 2 1 ° C for 15 minutes. 5 ml of the preculture broth was planted in this medium, and cultured at 37 ° C with shaking at 100 rpm for 48 hours to produce sorghum liquid mash. At this time, the enzyme activity of the liquid puppet, GA activity was 111.2 U / ml, and ASAA activity was 10.5 U / ml. -33- 200538545 3. Production method of rice shochu (1) Using yeast · Shochu yeast (Kagoshima yeast) (2) Brewing blending: The brewing blending is shown in Tables 丨 7 and 18. For rice, 90% milled rice is used. After the rice is washed, it is immersed for 15 minutes, drained for 10 minutes, and cooked for 30 minutes. The tests were divided into 2 test areas, 1) solid 麴 loading and brewing, 2) sorghum liquid 麴 loading and brewing, etc. The total rice and water content in the two test zones were equal. Yeast lines were cultured at 50 // 1 at 30 ° C for 48 hours and planted in YPD medium. (3) Fermentation conditions: 2 5 ° C (4) Distillation conditions: Decompression conditions (Table 17) Test area (sorghum liquid sorghum) Total first, second, and third stalks (g) 40.0 -11 40.0 Continue adding Rice (g) 311.3 507.6 507.6 1326.5 Liquid surface (m L) 500.0 _ 500.0 Water (mL) 594.0 765.4 265.6 1625.0 90% Lactic acid (mL) 1.4 — 1.4 (Table 1 8) Control area (solid radon) First The second and third times total indica rice (g) 311.3-a 311.3 Continue to add rice (g 1 _ 507.6 507.6 1015.2 water (mL) 594.0 765.4 765.6 2125.0 The solid tadpoles shown in Figure 8 and the control group are loaded into the brewing control. It can be clearly seen from the figure that compared with the control loading brewing using solid mash, the filling fermentation using sorghum liquid mash at -34- 200538545 has almost the same fermentation process. It is also loaded into the brewing area and sorghum. The alcohol content of the last liquor in liquid brewed into the brewing zone was about 19.1%. Secondly, the solid liquor was packed into the brewing zone and the sorghum liquid was packed into the brewing zone using the reduced pressure distillation method.醴 Steam retention, by 6 The professional judges used the 5-point evaluation method (good 1-3-5 difference) to perform the functional evaluation of the shochu raw wine prepared above. As a result, it was found that there was not much difference between the solid 麴 loading into the brewing zone and the sorghum liquid 麴 loading into the brewing zone. Sorghum liquid tincture can also produce shochu with sufficient quality. As shown in Table 19, the sorghum liquid tincture zone has a comment of "sweetness with grain" ^, showing that shochu that can be produced is significantly different from the previous solid tincture method Original wine. (Table 1 9) Functional evaluation results (average point) Wind rating solids 3.0 Gorgeous sorghum liquid 麴 3.0 Sweetness of cereals (Example 8) Production of liquid mash using corn (1) Pre-cultivation method: 8 g of 9 0% milled rice and 100m 1 water were put into a 500ml conical Erlenmeyer flask, placed in a high-temperature autoclave and sterilized at 121 ° C for 15 minutes. After cooling, the species of Aspergillus kawachii IFO4308 The spore planting spores were cultured in a pre-culture medium at a content of 1 × 10 6 cells / ml, and cultured at 37 ° C. with shaking at 100 rpm for 24 hours. (2) This culture method: 1 ~ 8g corn and 0.2g potassium nitrate, 0.3 g of potassium dihydrogen phosphate and 100 ml of water into 500 ml The Erlenmeyer flask with a plug is placed in an autoclave for 15 minutes at 121 ° C. 1 ml of the former culture medium is planted with 35-200538545 bacteria in this culture medium, and the temperature is 37 ° C to 100%. Incubate at rpm for 48 hours. The method of Experimental Example 1 was used to measure the amount of enzymes produced in the culture supernatant after culturing, that is, glucoamylase (GA) activity and acid-resistant α-amylase (ASA A) activity. The results are shown in Table 20. From Table 20, it is clear that the target glucoamylase of the enzyme activity required for the brewing of shochu is clearly 100 U / ml in the inspection area where the amount of corn is 4% or more. On the other hand, the target for acid-resistant α-amylase is iOu / m. As the amount of corn that cannot reach this target is increased, the amount of this enzyme tends to increase. As mentioned above, if the target of the AS AA activity in this test is unclear, because it has the ability to simultaneously generate GA enzymes and AS AA enzymes, it is very likely that the enzyme production will be increased to the target level by optimizing the culture conditions. . In addition, when shochu using 8% corn liquid mash is put into the brewing, if the ratio of mash is increased compared to ordinary blending, shochu can be sufficiently produced. (Table 20) Corn dosage enzyme activity (U / ml) GA ASAA 1 ° / 〇61.9 1.4 2% 66.4 1.7 4% 135.7 3.8 8% 114.2 4.2 (Example 9) Production of liquid mash using rough wheat (1) Cultivation method: 8 g of 65% milled wheat and 100 ml of water are placed in a 500 ml conical Erlenmeyer flask, placed in a high-temperature autoclave and sterilized at 12 1 ° C for 15 minutes. After cooling, the Aspergillus sp Shake for 24 hours. (2) The present culture method: Put 1 ~ 8g of brown wheat (95% milled wheat) and 0.2g of potassium nitrate, 0.3g of potassium dihydrogen phosphate and 100ml of water into a 500ml conical flask with a suppository, and put it in a high-temperature autoclave. The kettle was sterilized at 1 2 1 ° C for 15 minutes. 1 ml of the pre-culture broth was planted in this medium, and cultured at 37 ° C with shaking at 100 rpm for 48 hours. The method of Experimental Example 1 was used to measure the amount of enzyme produced in the culture supernatant after culturing, that is, glucoamylase (GA) activity and acid-resistant α-amylase (A S A A) activity. The results are shown in Table 21. It is clear from Table 21 that in the test area where the amount of rye is 4%, the target glucoamylase that knows the enzyme activity required for shochu brewing is clearly 100 U / ml and the acid-resistant α-amylase is clearly 1 OU / ml. From this, it can be seen that even if black fungus is used, it has the same high production effect of enzymes as white fungus. (Table 21) Rough wheat dosage Enzyme activity (U / ml) GA ASAA 1% 27.9 2.6 2% 41.5 2.0 4% 136.5 10.0 8 ° / 〇 7.1 0.1

(實施例1 0 )使用糙米（含粗殻米）的液體麴之製造 (1 )前培養方法：8g的90%碾製米（食米）和i〇〇ml 水放入5 00ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121 C進行15分鐘滅囷。冷卻後’將白麴囷（Aspergillus kawachii IF0 4308)的種麴孢子植菌於前培養基中，使含量 爲lx 106個/m卜並於371，以lOOrpm震盪培養24小時。 (2)本培養法：將1〜8g糙米（含粗殼）和〇.2g硝酸鉀、 -37- 200538545 0.3 g磷酸二氫鉀和1 0 0 m 1水放入5 0 0 m 1的附栓三角燒瓶；， 置於筒溫局壓滅菌签以1 2 1 °C進彳了 1 5分鐘滅菌。又，使用 的糙米係直接以含榖皮（粗殻）的狀態不經脫榖而被使用。 將1 ml前培養液植菌於此本培養基，並於37 °C，以1 〇〇rpm 震盪培養4 8小時。利用實驗例1的方法測定培養後的培養 上清液中的酵素生成量，亦即葡糖澱粉酶（GA )活性及耐 酸性α -澱粉酶（AS AA )活性。結果如表22所示。 從表22清楚可知，在糙米用量爲4%的試驗區，得知燒 酒釀造所需的酵素活性的目標値葡糖澱粉酶清楚爲 ® 100U/ml及耐酸性α -澱粉酶清楚爲10U/ml。由此可知，即 使使用含榖皮（粗殼）的米仍具有和糙麥相同的酵素的高 生成效果。 (表 22 ) 糙米（含粗殼米）用量 酵素活性（U/ml) GA ASAA 1°/〇 29.2 0.8 2% 39.3 1.8 4% 140.4 11.5 8% 89.0 5.2 (實施例1 1 )使用糙米（含粗殼米）的液體麴之製造(Example 10) Production of liquid rice using brown rice (containing coarse-shelled rice) (1) Pre-cultivation method: 8 g of 90% milled rice (rice) and 100 ml of water were put into a 500 ml attachment plug Erlenmeyer flask, placed in an autoclave at 121 C for 15 minutes for extinction. After cooling, the seed spores of Aspergillus kawachii IF0 4308 were planted in a pre-culture medium at a content of 1 x 106 cells / m2 and cultured at 371 at 100 rpm for 24 hours with shaking. (2) This culture method: Put 1 ~ 8g of brown rice (including coarse shell) and 0.2g of potassium nitrate, -37- 200538545 0.3 g of potassium dihydrogen phosphate and 100 m 1 of water into a 500 m 1 The Erlenmeyer flask was plugged; the tube was placed under a local pressure autoclave and sterilized at 121 ° C for 15 minutes. In addition, the used brown rice is used in a state containing coriander skin (coarse husk) without being removed. 1 ml of pre-culture broth was planted in this medium, and cultured at 37 ° C with shaking at 1000 rpm for 48 hours. The method of Experimental Example 1 was used to measure the amount of enzyme produced in the culture supernatant after culture, that is, glucoamylase (GA) activity and acid alpha-amylase (AS AA) activity. The results are shown in Table 22. It is clear from Table 22 that in the test area where the amount of brown rice is 4%, the target of the enzyme activity required for shochu brewing is clear. The glucoamylase is clearly 100 U / ml and the acid-resistant α-amylase is clearly 10 U / ml. . From this, we can see that even when rice containing coriander skin (coarse husk) is used, it has the same high enzyme production effect as that of rye. (Table 22) Enzyme activity (U / ml) of brown rice (including coarse-hulled rice) GA ASAA 1 ° / 〇29.2 0.8 2% 39.3 1.8 4% 140.4 11.5 8% 89.0 5.2 (Example 1 1) Shell m) manufacturing of liquid tincture

(1 )前培養方法：的90%碾製米（食米）和100ml 水放入5 00ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121 °C進行15分鐘滅菌。冷卻後，將泡盛麴菌（Aspergillus awamori IF04388)的種麹孢子植菌於前培養基中，使含量 爲lxlO6個/m卜並於37°C，以lOOrpm震盪培養24小時。 (2 )本培養法：將1〜8 g縫米（含粗殼）和〇 · 2 g硝酸鉀、 -38- 200538545 0.3g磷酸二氫鉀和l〇〇ml水放入500ml的附栓三角燒瓶， 置於高溫高壓滅菌釜以1 2 1 °C進行1 5分鐘滅菌。又，使用 的糙米係直接以含榖皮（粗殼）的狀態不經脫榖而被使用。 將lml前培養液植菌於此本培養基，並於37°C，以10〇rpm 震盪培養4 8小時。利用實驗例1的方法測定培養後的培養 上清液中的酵素生成量，亦即葡糖澱粉酶（GA )活性及耐 酸性α -澱粉酶（AS AA )活性。結果如表23所示。 從表23清楚可知，在糙米用量爲8%的試驗區，得知燒 酒釀造所需的酵素活性的目標値葡糖澱粉酶清楚爲 ® 100U/ml及耐酸性α -澱粉酶清楚爲10U/ml。由此可知，使 用含榖皮（粗殼）的米且使用泡盛麴菌，仍具有酵素的高 生成效果。 (表 2 3 ) 糙米（含粗殼米）用量 酵素活性（U/ml) GA ASAA 1°/〇 14.3 1.4 2% 19.3 1.4 4% 40.3 3.9 8% 100.0 10.5(1) Pre-cultivation method: 90% of milled rice (rice) and 100 ml of water are put into a 500 ml conical Erlenmeyer flask, and placed in a high-temperature autoclave at 121 ° C for 15 minutes for sterilization. After cooling, the Aspergillus awamori IF04388 spore-forming spores were planted in the pre-culture medium at a content of 1 × 10 6 cells / m and cultured at 37 ° C. with shaking at 100 rpm for 24 hours. (2) This culture method: Put 1 ~ 8 g seam rice (including coarse shell) and 0.2 g potassium nitrate, -38- 200538545 0.3 g potassium dihydrogen phosphate and 100 ml water into 500 ml triangle with plug The flask was placed in a high-temperature autoclave and sterilized at 12 1 ° C for 15 minutes. In addition, the used brown rice is used in a state containing coriander skin (coarse husk) without being removed. 1 ml of the pre-culture broth was planted in this medium, and cultured at 37 ° C with shaking at 100 rpm for 4 8 hours. The method of Experimental Example 1 was used to measure the amount of enzyme produced in the culture supernatant after culture, that is, glucoamylase (GA) activity and acid alpha-amylase (AS AA) activity. The results are shown in Table 23. From Table 23, it is clear that in the test area where the amount of brown rice is 8%, the target of the enzyme activity required for the brewing of shochu is clear. The glucoamylase is clearly 100 U / ml and the acid-resistant α-amylase is clearly 10 U / ml. . From this, we can see that using rice containing coriander husk (coarse shell) and using awamori fungus still has a high enzyme production effect. (Table 2 3) Consumption of brown rice (including coarse shell rice) Enzyme activity (U / ml) GA ASAA 1 ° / 〇 14.3 1.4 2% 19.3 1.4 4% 40.3 3.9 8% 100.0 10.5

(實施例1 2 )使用去除粗殼的糙米的液體麴之製造 1 .前培養方法： 的90%碾製米和100ml水放入5 00ml的附栓三角燒 瓶，置於高溫高壓滅菌爸以1 2 1 °C進行I 5分鐘滅菌。冷卻 後，將白麹菌（Aspergillus kawachii IF0 4308)的種麴孢子 植菌於前培養基中，使含量爲lxlO6個/m卜並於37°C，以 1 0 0 r p m震盪培養2 4小時。 -39- 200538545 2.本培養法： 將1〜l〇g去除粗殼的糙米和〇.2g硝酸鉀、〇.3g磷酸二氫 鉀和100ml水放入5 00ml的附栓三角燒瓶，置於高溫高壓 滅菌釜以1 2 1°C進行1 5分鐘滅菌。冷卻後將1 ml前培養液 植菌於此本培養基，並於37t，以lOOrpm震盪培養48小 時。 另一方面，對照組係將1〜l〇g的90%碾製米和〇.2g硝 酸鉀、〇.3g磷酸二氫鉀和100ml水放入500ml的附栓三角 燒瓶，置於高溫高壓滅菌釜以1 2 1°C進行1 5分鐘滅菌。冷 Φ 卻後將1ml前培養液植菌於此本培養基，並於37t，以 lOOrpm震盪培養48小時。 培養結束後，測定各培養上清液中的葡糖澱粉酶活性 (GA )和耐酸性α -澱粉酶活性（ASAA )。使用糖化力分 別定量套組（龜甲萬製）測定葡糖澱粉酶活性（GA )，耐 酸性α -澱粉酶活性（ASAA )之測定方法，係將（非專利 文獻3 )的方法稍加改良，培養物經過酸處理後使非耐酸 性α -澱粉酶失活後，使用α澱粉酶測定套組（龜甲萬製） φ 進行測定。更具體而言，在1ml培養液中添加9ml的100mM 醋酸緩衝液（pH3 )，於37°C進行1小時酸處理後，使用 α澱粉酶測定套組（龜甲萬製）進行測定。 3 .結果： 如表24所示。至目前爲止的硏究，釀造燒酒所需的酵 素活性，葡糖澱粉酶爲l〇〇U/ml、耐酸性α-澱粉酶爲10U/ml 即可。對照組使用白米時，GA及ASAA無法同時超越目標 値，而試驗組使用糙米時，GA及ASAA可同時平衡地生成， -40- 200538545 特別是糙米用量8 %以上時’目標酵素活性値變清楚。根 據此結果，糙米較白米更適合作爲液體麴之原料。 (表 24 ) 試驗組 對照組 糙米用量 酵素活性（U/ml) 白米用量 酵素活性（U/ml) GA ASAA GA ASAA 1% 52.9 2.6 1°/〇 19.8 1.1 2% 64.0 4.0 2¾ 31.9 2.0 4% 101.8 9.3 4°/〇 51.3 4.3 8% 112.7 15.6 8% 56.5 10.3 1〇°/〇 136.2 18.7 10% 52.1 11.2 (實施例1 3 )以使用去除粗殼的糙米的液體麹製造燒酒 1.固體麴製造方法 使用90%碾製米，洗米後浸漬15分鐘，瀝乾10分鐘， 並蒸煮30分鐘後使冷卻至40 °C，對於每lkg碾製米，植菌 lg 的種麴（白麹菌 Aspergillus kawachii IF0 4308)，於 40 t ·相對溼度95%的條件培養6小時，再於30 °C ·相對溼 度90%下培養18小時。 2.液體麴之製法 (1 )前培養方法·· 8g的90%碾製米和100ml水放入 5 0 0ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121 °C進行 15分鐘滅菌。冷卻後，將白麹菌（Aspergillus kawachii IFO4308 )的種麴孢子植菌於前培養基中，使含量爲lxl〇6 個/ml，並於37°C，以l〇〇rpm震盪培養24小時。 (2 )本培養法：將40g去除粗殼的糙米和l.Og硝酸鉀、 1.5g磷酸二氫鉀和5 00ml水放入2000ml的附栓三角燒瓶， -41- 200538545 置於高溫高壓滅菌釜以1 2 1 °C進行1 5分鐘滅菌。將5 m 1前 培養液植菌於此本培養基，並於37 °C，以100rpm震盪培養 4 8小時，製造糙米液體麴。 對照組係將4〇2的90%碾製米和1.〇2硝酸鉀、1.52磷酸 二氫紳和500ml水放入2000ml的附栓三角燒瓶，置於高溫 高壓滅菌釜以1 2 1 °C進行1 5分鐘滅菌。將5 ml前培養液植 菌於此本培養基，並於37°C，以lOOrpm震盪培養48小時， 製造白米液體麴。 3.米燒酒之製法 (1) 使用酵母：燒酒酵母（鹿兒島酵母） (2) 釀造掺合：釀造掺合如表25〜表27所示。米係使 用9 0%碾製米，洗米後浸漬15分鐘，瀝乾10分鐘，並蒸 煮3 0分鐘後的米。實驗區（試驗區、對照區）分爲1 )固 體麴裝入釀造、2)糙米液體麴裝入釀造、及3)白米液體 麵裝入釀造等3區’各區的總米量及水量相等。酵母係將 5 0// 1於3CTC靜置培養48小時者植菌於ypd培養基。 (3 )發酵條件：25t：，固定 (4 )蒸留條件：減壓蒸留(Example 1 2) Manufacture of liquid mash using brown rice with coarse shell removed 1. Pre-cultivation method: 90% milled rice and 100 ml of water were placed in a 500 ml conical Erlenmeyer flask and placed in a high-temperature autoclave Sterilize at 21 ° C for 1 minute. After cooling, the seed spores of Aspergillus kawachii IF0 4308 were planted in the pre-culture medium at a content of 1 × 10 6 cells / m at 37 ° C and shaken at 100 r p m for 2 4 hours. -39- 200538545 2. This culture method: Put 1 ~ 10g of brown rice with coarse shells removed, 0.2g of potassium nitrate, 0.3g of potassium dihydrogen phosphate, and 100ml of water into a 500ml conical Erlenmeyer flask and place The autoclave was sterilized at 12 1 ° C for 15 minutes. After cooling, 1 ml of the pre-culture broth was planted in this medium, and cultured at 37 t with shaking at 100 rpm for 48 hours. On the other hand, in the control group, 1 to 10 g of 90% milled rice and 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate, and 100 ml of water were placed in a 500 ml conical Erlenmeyer flask and placed in an autoclave. The kettle was sterilized at 1 2 1 ° C for 15 minutes. After cold Φ, 1ml of the former culture liquid was planted in this medium, and cultured at 37t with shaking at 100 rpm for 48 hours. After the culture was completed, glucoamylase activity (GA) and acid-resistant α-amylase activity (ASAA) in each culture supernatant were measured. The method for measuring glucoamylase activity (GA) and acid-resistant α-amylase activity (ASAA) by using a saccharification power quantitative set (made by Kikkoman) is a slightly improved method (Non-Patent Document 3). After the culture was acid-treated to inactivate non-acid-resistant α-amylase, the α-amylase measurement kit (made by Kikkoman) was used to measure φ. More specifically, 9 ml of 100 mM acetate buffer (pH 3) was added to 1 ml of the culture solution, and after acid treatment at 37 ° C for 1 hour, the measurement was performed using an α-amylase measurement kit (manufactured by Kikkoman). 3. Results: As shown in Table 24. So far, the enzyme activity required for brewing shochu is 100 U / ml for glucoamylase and 10 U / ml for acid-resistant α-amylase. When using white rice in the control group, GA and ASAA cannot exceed the target 値 at the same time, and when using brown rice in the test group, GA and ASAA can be produced at the same time in a balanced manner. -40- 200538545 Especially when the amount of brown rice is more than 8%, the target enzyme activity 値 becomes clear. . Based on this result, brown rice is more suitable as a raw material for liquid rice than white rice. (Table 24) Enzyme activity of brown rice dosage (U / ml) Enzyme activity of white rice dosage (U / ml) GA ASAA GA ASAA 1% 52.9 2.6 1 ° / 〇19.8 1.1 2% 64.0 4.0 2¾ 31.9 2.0 4% 101.8 9.3 4 ° / 〇51.3 4.3 8% 112.7 15.6 8% 56.5 10.3 10 ° / 〇136.2 18.7 10% 52.1 11.2 (Example 1 3) Production of shochu using liquid rice cake with rough rice removed 1. Solid rice cake manufacturing method Use 90% milled rice, soak the rice for 15 minutes, drain for 10 minutes, and cook for 30 minutes to cool to 40 ° C. For every 1 kg of milled rice, plant lg seed worm (Aspergillus kawachii IF0 4308) ), Incubate at 40 t · 95% relative humidity for 6 hours, and then at 30 ° C · 90% relative humidity for 18 hours. 2. Production method of liquid mash (1) Pre-cultivation method ... 8g of 90% milled rice and 100ml of water are placed in a 500ml conical flask with a stopper and placed in a high-temperature autoclave at 121 ° C for 15 minutes for sterilization. After cooling, Aspergillus kawachii IFO4308 was seeded in the pre-culture medium to a content of 1 × 106 cells / ml, and cultured at 37 ° C. with shaking at 100 rpm for 24 hours. (2) The culture method: Put 40 g of brown rice with coarse shells removed and 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate and 500 ml of water into a 2000 ml conical Erlenmeyer flask, -41- 200538545 placed in a high-temperature autoclave Sterilize at 1 2 1 ° C for 15 minutes. 5 m 1 of the pre-culture liquid was planted in this medium, and cultured at 37 ° C with shaking at 100 rpm for 4 8 hours to produce a brown rice liquid tincture. In the control group, 90% milled rice of 4.02, potassium nitrate 1.02, dihydrogen phosphate 1.52, and 500 ml of water were placed in a 2000 ml Erlenmeyer flask with a plug and placed in an autoclave at 1 2 ° C. Sterilize for 15 minutes. 5 ml of pre-culture broth was planted in this medium, and cultured at 37 ° C. with shaking at 100 rpm for 48 hours to produce a white rice liquid tincture. 3. Method for making rice shochu (1) Using yeast: Shochu yeast (Kagoshima yeast) (2) Brewing blend: Brewing blends are shown in Table 25 to Table 27. Rice is 90% milled rice, washed for 15 minutes, soaked for 10 minutes, and cooked for 30 minutes. The experimental area (experimental area, control area) is divided into 1) solid 麴 into the brewing, 2) brown rice liquid 酿 into the brewing, and 3) white rice liquid noodle into the brewing, etc. The total amount of rice and water in each area are equal . Yeast lines were cultured for 50 hours at 1/1/1 at 3CTC for 48 hours, and then planted in ypd medium. (3) Fermentation conditions: 25t :, fixed (4) Distillation conditions: reduced pressure distillation

(表 25 ) 試驗區（糙米液體麹） 第一次 第二次 第三次 合計 麴米（g) 40.0 一 一 40.0 續加入米（g) 271.3 507.6 507.6 1286.5 水（mL· ) 594.0 765.4 265.6 1625.0 液體麴（mL) 500.0 一 一 500.0 90% 乳酸（mL) 1.4 — 一 1.4 -42- 200538545 (表 26 ) 對照區（1)(白米液體麴） 第一次 第二次 第三次 合計 麴米（g) 40.0 — — 40.0 續加入米（g) 271.3 507.6 507.6 1286.5 水（m L ) 594.0 765.4 265.6 1625.0 液體麹（mL) 500.0 一 一 500.0 90% 乳酸（mL) 1.4 — — 1.4 (表 27) 對照區（2 )(固體麴） 第一次 第二次 第三次 合計 麴米（g ) 311.3 一 — 311.3 續加入米（g ) — 507.6 507.6 1015.2 水（mL ) 594.0 765.4 765.6 2125.0 4.結果與硏究 發酵過程如第9圖所示。從圖清楚可知，固體麴裝入釀 造區和糙米液體麹裝入釀造區，具有幾乎相同的發酵過 程。惟，白米液體麹裝入釀造區的發酵經過較差。又’最 • 後酒醴的酒精度數，固體麴裝入釀造區19.1%和糙米液體 麴裝入釀造區18.9%幾乎相同程度，相對於此，白米液體 麹裝入釀造區的最後酒醴的酒精度數爲12.5，明顯地較前 二者低。 以減壓蒸留法使製得固體麴裝入釀造區和糙米液體麴 裝入釀造區的燒酒酒醴進行蒸留，由6位專業品評員以5 點品評法（佳1-3-5差）進行上述製得的燒酒原酒之官能品 評，結果如表2 8所示，可知固體麴裝入釀造區和糙米液體 -43 - 200538545 麹裝入釀造區無太大的差異。惟，糙米液體麴區有「舒暢 •輕快」之評語，可製得具舒暢香味的米燒酒。從上述的 結果顯示，即使使用糙米液體麴亦可製造和固體麴相同品 質的米燒酒。 (表 28) 官能品評結果 評點（平均點） 風評 固體麴 3.0 華麗 糙米液體麴 2.8 華麗、輕快、舒暢(Table 25) Test area (Brown rice liquid rice) Total rice rice (g) for the first time, second time, and third time (g) 40.0-10.0 Continue to add rice (g) 271.3 507.6 507.6 1286.5 Water (mL ·) 594.0 765.4 265.6 1625.0 Liquid rice ( mL) 500.0-500.0 90% lactic acid (mL) 1.4 —-1.4 -42- 200538545 (Table 26) Control area (1) (white rice liquid tincture) Total rice grain (g) 40.0 — — 40.0 Continue to add rice (g) 271.3 507.6 507.6 1286.5 Water (m L) 594.0 765.4 265.6 1625.0 Liquid tritium (mL) 500.0-500.0 90% Lactic acid (mL) 1.4 — 1.4 (Table 27) Control area (2) (solid麴) The first, second, and third times total indica rice (g) 311.3 One — 311.3 Continue to add rice (g) — 507.6 507.6 1015.2 Water (mL) 594.0 765.4 765.6 2125.0 4. Results and research fermentation process as shown in Figure 9 Show. It is clear from the figure that the solid 麴 loaded into the brewing zone and the brown rice liquid 麹 loaded into the brewing zone have almost the same fermentation process. However, the fermentation of the white rice liquid mash into the brewing area was poor. Also, the alcohol content of the last wine 醴 is almost the same as that of solid rice 1 19.1% in the brewing zone and the brown rice liquid 1 18.9% in the brewing zone. In contrast, the white rice liquid 麹 is loaded into the alcohol of the last wine 区 in the brewing zone. The degree is 12.5, which is significantly lower than the former two. The prepared solid 麴 was placed in the brewing area and brown rice liquid 麴 into the shochu wine 酿 in the brewing area by the reduced-pressure steaming method. The steaming was performed by 6 professional judges using a 5-point evaluation method (good 1-3-5 difference). The functional evaluation of the shochu original liquor prepared as described above is shown in Table 2-8. It can be seen that there is not much difference between the solid 麴 loading in the brewing zone and the brown rice liquid -43-200538545 麹 loading into the brewing zone. However, there is a comment on "smoothing • brisk" in the liquid rice area of brown rice, which can make rice shochu with a smooth flavor. From the above results, it is shown that even when using brown rice liquid, it is possible to produce rice shochu of the same quality as solid rice. (Table 28) Functional evaluation results Evaluation point (average point) Wind evaluation Solid 麴 3.0 Gorgeous Brown rice liquid 麴 2.8 Gorgeous, light and comfortable

(實施例1 4 )使用去除粗殻的糙米的液體麴之製法 1.前培養方法： 8g的90%碾製米（食米）和l〇〇ml水放入500ml的附栓 三角燒瓶，置於高溫高壓滅菌釜以1 2 KC進行1 5分鐘滅 菌。冷卻後，將泡盛麹菌（Aspergillus awamori IF04388) 的種麴孢子植菌於前培養基中，使含量爲lxl〇6個/m卜並 於37°C，以100ι·ριη震盪培養24小時。 2.本培養法： 將1〜8g去除粗殻的糙米和〇.2g硝酸鉀、0.3g磷酸二氫 鉀和1 0 0 m 1水放入5 0 0 m 1的附栓三角燒瓶，置於高溫高壓 滅菌釜以1 2 1 °C進行1 5分鐘滅菌。冷卻後將1 m 1前培養液 植菌於此本培養基，並於37°C，以lOOrpm震盪培養48小 培養結束後，以實施例1 2的方法測定培養上清液中的 葡糖澱粉酶活性（GA )和耐酸性α -澱粉酶活性（AS Aa )。 3.結果： 如表29所示。如前述般，釀造燒酒所需的酵素活性的 -44- 200538545 目標値’葡糖澱粉酶爲l〇OU/ml、耐酸性α -澱粉酶爲 10U/ml。從表29清楚可知，在糙米用量爲8%的試驗區， G A和AS A A的目標値均清楚，即使使用泡盛麴菌亦可達到 和白麹菌相同的酵素高生成效果。又，更增加糙米的用量 時’亦可期待產生和白麴菌相同的酵素高生成效果。 (表 29 ) 糙米用量 酵素活性（U/ml) GA ASAA 1% 23.0 0.1 2% 33.0 0.2 4% 89.0 8.2 8% 132.7 10.6(Example 1 4) Preparation method of liquid rice using coarse rice and brown rice removed 1. Pre-cultivation method: 8 g of 90% milled rice (rice) and 100 ml of water were placed in a 500 ml Erlenmeyer flask with a stopper. Sterilize in a high-temperature autoclave at 12 KC for 15 minutes. After cooling, Aspergillus awamori IF04388 spore-forming spores were planted in the pre-culture medium at a content of 1 × 106 / m and cultured at 37 ° C. with shaking at 100 μm for 24 hours. 2. This culture method: Put 1 ~ 8g of brown rice with coarse shell removed, 0.2g of potassium nitrate, 0.3g of potassium dihydrogen phosphate, and 100m 1 of water into a 500 m 1 conical flask with a stopper and place The autoclave was sterilized at 121 ° C for 15 minutes. After cooling, 1 m 1 of the pre-culture broth was planted in this medium, and the culture was performed at 37 ° C with shaking at 100 rpm for 48 hours. After the completion of the culture, the glucoamylase in the culture supernatant was measured by the method of Example 12 Activity (GA) and acid-resistant α-amylase activity (AS Aa). 3. Results: As shown in Table 29. As mentioned above, the target enzyme activity required for brewing shochu -44-200538545 is 10 OU / ml, and the acid-resistant α-amylase is 10 U / ml. It is clear from Table 29 that in the test area where the amount of brown rice was 8%, the target slugs of G A and AS A A were clear, and the same high enzyme production effect as that of white slugs could be achieved even with Awamori slugs. When the amount of brown rice is further increased, it is also expected to produce the same high enzyme production effect as that of white fungus. (Table 29) Brown rice dosage Enzyme activity (U / ml) GA ASAA 1% 23.0 0.1 2% 33.0 0.2 4% 89.0 8.2 8% 132.7 10.6

(實施例1 5 )大豆液體麹之製法 1.前培養方法： 8g的90%碾製米和100ml水放入500ml的附栓三角燒 瓶，置於高溫高壓滅菌釜以1 2 1 °C進行1 5分鐘滅菌。冷卻 後，將白麴菌（Aspergillus kawachii IF 0 4308)的種麴孢子 植菌於前培養基中，使含量爲lxl 06個/m卜並於37 °C，以 1 0 0 r p m震盪培養2 4小時。 2.本培養法： 將1〜10g大豆和〇.2g硝酸鉀、0.3g磷酸二氫鉀和100ml 水放入5 00ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121 t進行1 5分鐘滅菌。冷卻後將1 ml前培養液植菌於此本培 養基，並於3 7 °C，以1 〇 〇 rp m震盪培養4 8小時。培養結束 後，測定培養上清液中的葡糖澱粉酶（G A )活性和耐酸性 α -澱粉酶（AS AA )活性。使用糖化力分別定量套組（龜 -45- 200538545 甲萬製）測定GA活性，ASAA活性之測定方法，係將 專利文獻3 )的方法稍加改良，培養物經過酸處理後 耐酸性α -澱粉酶失活後，使用α —澱粉酶測定套組（ 萬製）進行測定。更具體而言，在1 ml培養液中添力口 的100mM醋酸緩衝液（ph3 )，於37°C進行1小時酸 後’使用α -澱粉酶測定套組（龜甲萬製）進行測定。 3.結果： 如表30所示。至目前爲止的硏究，釀造燒酒所需 素活性的目標値，葡糖澱粉酶爲l〇〇U/ml、耐酸性α -酶爲10U/ml。從表清楚可知，隨著大豆用量的增加 活性及AS AA活性亦增加，大豆用量8%以上時，目標 活性値變清楚。根據此結果，大豆適合作爲液體麴之原 (表 30 ) 大豆用量 酵素活性（U/ml) GA ASAA 1% 43.6 2.3 2% 48.0 2.7 4% 84.5 9.3 8% 110.8 11.3 10% 103.7 10.8 (實施例1 6 )以使用大豆的液體麴製造米燒酒 1 .固體麴製造方法 使用90%碾製米，洗米後浸漬15分鐘，瀝乾10夕 並蒸煮30分鐘後使冷卻至40 °C，對於每lkg碾製米， lg 的種麴（白麹菌 Aspergillus kawachii IF0 4308)， °C ·相對溼度95%的條件培養24小時，又於35°C · (非 使非 龜甲 9ml 處理 的酵 澱粉 ，GA 酵素 料0 、鐘， 植菌 於40 相對 -46 - 200538545 溼度9 5 %下培養6小時，再於3 0 °C ·相對溼度9 0 %下培養 1 8小時。 2. 液體麴之製法 (1 )前培養方法： 8g的90%碾製米和1 〇〇ml水放入500ml的附栓三角燒 瓶，置於高溫高壓滅菌釜以1 2 1 °C進行1 5分鐘滅菌。冷卻 後，將白麴菌（Aspergillus kawachii IF0 4308)的種麴孢子 植菌於前培養基中，使含量爲lxl〇6個/m卜並於37°C，以 1 0 0 r p m震盪培養2 4小時。 (2 )本培養法： 將40g大豆和l.Og硝酸鉀、1.5g磷酸二氫鉀和500ml 水放入 2000ml的附栓三角燒瓶，置於高溫高壓滅菌釜以 1 2厂C進行1 5分鐘滅菌。將5 ml前培養液植菌於此本培養 基，並於37°C，以lOOrpm震盪培養48小時，製造大豆液 體麴。 3. 米燒酒之製法 (1 )使用酵母··燒酒酵母（鹿兒島酵母） • ( 2 )釀造掺合： 釀造掺合如表31、表32所示。米係使用90%碾製米’ 洗米後浸漬1 5分鐘，瀝乾1 0分鐘，並蒸煮30分鐘後的米。 試驗區分爲1)固體麹裝入釀造、2)大豆液體麴裝入釀造 等2試驗區，二試驗區的總米量及水量相等。酵母係將5 0 // 1於30°C靜置培養48小時者植菌於YPD培養基。 (3 )發酵條件·· 2 5 °C，固定 (4 )蒸留條件：減壓蒸留 -47- 200538545 (表 31 ) -----Ί 試驗區（大豆液體麴） 第一次 第二次 第三次 ------ 合計 麴大豆（g) 40.0 一 — 40.0 續加入米（g) 311.3 507.6 507.6 1 326.5 水（m L ) 594.0 765.4 265.6 1625.0 液體麹（mL) 500.0 — 一 500.0 90% 乳酸（mL) 1.4 — — 1.4 (表 32) 對照區（ 固體麴） 第一次 第二次 第三次 合計 麹米（g) 311.3 一 一 311.3 續加入米（g ) 一 507.6 507.6 1015.2 水（m L ) 594.0 765.4 765.6 2125.0 4.結果與硏究(Example 15) Method for preparing soybean liquid tincture 1. Pre-cultivation method: 8g of 90% milled rice and 100ml of water were placed in a 500ml Erlenmeyer flask with a plug, and placed in a high-temperature autoclave at 1 2 1 ° C for 1 5 minutes sterilization. After cooling, the spores of Aspergillus kawachii IF 0 4308 were planted in the pre-culture medium at a content of lxl 06 cells / m at 37 ° C and shaken at 100 rpm for 2 4 hours. . 2. This culture method: Put 1 ~ 10g of soybean and 0.2g of potassium nitrate, 0.3g of potassium dihydrogen phosphate and 100ml of water into a 500ml conical Erlenmeyer flask, and place in a high-temperature autoclave at 121 t for 15 minutes Sterilize. After cooling, 1 ml of the pre-culture broth was planted in this medium, and cultured at 37 ° C for 4 8 hours with shaking at 1000 rpm. After the completion of the culture, the glucoamylase (GA) activity and the acid-resistant α-amylase (AS AA) activity in the culture supernatant were measured. The saccharification power was used to quantify the GA activity (Kame-45- 200538545 by Kowan) to determine GA activity, and the method for measuring ASAA activity was slightly improved. The culture was acid-treated and treated with acid-resistant α-starch. After the enzyme was inactivated, the measurement was performed using an α-amylase assay kit (10,000 system). More specifically, 100 ml of acetic acid buffer solution (ph3) was added to 1 ml of the culture solution, and acidified at 37 ° C for 1 hour. The measurement was performed using an α-amylase measurement kit (manufactured by Kikkoman). 3. Results: As shown in Table 30. So far, the target of the activity required for brewing shochu is 100 U / ml for glucoamylase and 10 U / ml for acid-resistant α-enzyme. It is clear from the table that the activity and AS AA activity also increase with the increase in the amount of soybeans. When the amount of soybeans exceeds 8%, the target activity becomes clear. Based on this result, soybean is suitable as the source of liquid mash (Table 30). Soybean dosage Enzyme activity (U / ml) GA ASAA 1% 43.6 2.3 2% 48.0 2.7 4% 84.5 9.3 8% 110.8 11.3 10% 103.7 10.8 (Example 1 6) Production of rice shochu using liquid soy using soybeans 1. The method for producing solid soy uses 90% milled rice, soak the rice for 15 minutes, drain it for 10 minutes, and cook for 30 minutes to cool to 40 ° C. For each 1 kg of rice Rice making, lg seed cultivar (Aspergillus kawachii IF0 4308), ° C · 95% relative humidity for 24 hours, and 35 ° C · (non-tortoise 9ml-treated fermented starch, GA enzyme material 0 , Bell, and plant at 40 relative -46-200538545 humidity of 95% for 6 hours, and then at 30 ° C · relative humidity of 90% for 18 hours. 2. culture method of liquid tincture (1) before cultivation Method: 8g of 90% milled rice and 100ml of water were put into a 500ml Erlenmeyer flask with a plug and placed in a high-temperature autoclave for 15 minutes at 121 ° C. After cooling, the white fungus ( Aspergillus kawachii IF0 4308) was planted in the pre-culture medium to a content of 1 × 106 / m The culture was incubated at 37 ° C with shaking at 100 rpm for 24 hours. (2) This culture method: 40 g of soybeans and 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate and 500 ml of water were put into a 2000 ml of suppository The conical flask was placed in a high-temperature autoclave and sterilized for 15 minutes at 12 Plant C. 5 ml of the pre-culture broth was planted in this medium, and cultured at 37 ° C with shaking at 100 rpm for 48 hours to produce a soybean liquid tincture. 3. Method for making rice shochu (1) using yeast ·· shochu yeast (Kagoshima yeast) • (2) brewing blending: The brewing blending is shown in Table 31 and Table 32. The rice system uses 90% milled rice ' After washing the rice, soak it for 15 minutes, drain it for 10 minutes, and cook the rice after 30 minutes. The test is divided into 1) solid 麹 into the brewing, 2) soybean liquid 麴 into the brewing, etc. 2 test areas, the total of the two test areas The amount of rice and water is equal. Yeast strains were cultured at 30 ° C for 48 hours and planted in YPD medium. (3) Fermentation conditions · 2 5 ° C, fixed (4) Distillation conditions: reduced pressure distillation -47- 200538545 (Table 31) ----- Ί Test area (soy liquid 麴) First time Second time Third time ------ Total loquat soybeans (g) 40.0 I-40.0 Continue to add rice (g) 311.3 507.6 507.6 1 326.5 Water (m L) 594.0 765.4 265.6 1625.0 Liquid tritium (mL) 500.0 — One 500.0 90% lactic acid (mL) ) 1.4 — — 1.4 (Table 32) Control area (solid radon) Total rice (g) for the first, second and third times 311.3-311.3 Continue to add rice (g)-507.6 507.6 1015.2 Water (m L) 594.0 765.4 765.6 2125.0 4. Results and research

發酵過程如第1 〇圖所示。從圖清楚可知，對照區的米 固體_裝入釀造區和試驗區的大豆液體麴裝入釀造區，具 有幾乎相同的發酵過程。又，最後酒醴的酒精度數，米固 體麴裝入釀造區19.1% 、大豆液體麴裝入釀造區18·7%幾 乎相同程度。 以減壓蒸留法使製得米固體麴裝入釀造區、大豆液體麴 裝入釀造區的燒酒酒醴進行蒸留，由6位專業品評員以5 點品評法（佳1 - 3 - 5差）進行上述製得的燒酒原酒之官能品 評’結果如表3 3所示，可知米固體麴裝入釀造區、大豆液 體麴裝入釀造區無太大的差異，可知大豆液體麴亦可製造 -48- 200538545 具充分品質的燒酒。又，大豆液體麴區有「華麗的香味」 之評語’顯示可製造和先前固體麴製法有明顯差異化之燒 酒原酒。 (表 33 ) 官能品評結果 評點（平均點） 風評 固體麴 3.0 舒暢 大豆液體麴 2.6 華麗、輕快 (實施例1 7 )紅豆液體麴之製法 1 ·前培養方法： 8g的90%碾製米和1〇〇ml水放入500ml的附栓三角燒 瓶’置於高溫高壓滅菌釜以1 2 1 °C進行1 5分鐘滅菌。冷卻 後’將白麹菌（Aspergillus kawachii IFO4308)的種麴孢子 植菌於前培養基中，使含量爲lxl〇6個/m卜並於37t，以 lOOrpm震盪培養24小時。 2. 本培養法： 將1〜l〇g紅豆和〇.2g硝酸鉀、0.3g磷酸二氫鉀和100ml φ 水放入500ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121 °C進行1 5分鐘滅菌。將1 ml前培養液植菌於此本培養基， 並於3 7 C，以1 〇 〇 r p m震盪培養4 8小時。培養結束後，以 實施例1 5的方法測定培養上清液中的葡糖澱粉酶（g A ) 活性和耐酸性α -澱粉酶（A S A A )活性。 3. 結果：The fermentation process is shown in Figure 10. It is clear from the figure that the rice solids in the control zone are loaded into the brewing zone and the soybean liquid tincture in the test zone has almost the same fermentation process. In addition, the final alcohol content of rice wine was approximately 19.1% for rice solids and 18.7% for soybean liquids. The solid rice 麴 produced in the brewing zone and the soy liquid 麴 were placed in the shochu wine 醴 in the brewing zone by the reduced-pressure evaporation method. The 6 professional judges evaluated the method by 5 points (good 1-3-5 difference) The results of the functional evaluation of the shochu raw liquor prepared above are shown in Table 33. It can be seen that there is not much difference between rice solid 麴 loaded into the brewing zone and soybean liquid 麴 loaded into the brewing zone, and it can be seen that soybean liquid 麴 can also be produced -48 -200538545 Shochu with full quality. In addition, the review of the "grand scent" in the soy liquid region shows that it is possible to produce shochu original sake that is significantly different from the previous solid method. (Table 33) Functional evaluation result evaluation points (average point) Wind evaluation solid 麴 3.0 Soothing soy liquid 麴 2.6 Gorgeous and brisk (Example 17) Method 1 for making red bean liquid · Pre-cultivation method: 8g of 90% milled rice and 1 〇OOml of water was put into a 500ml Erlenmeyer flask with a stopper and placed in a high-temperature autoclave for sterilization at 12 1 ° C for 15 minutes. After cooling, the seed spores of Aspergillus kawachii IFO4308 were planted in a pre-culture medium at a content of 1 × 106 cells / m2 and cultured at 37 t with shaking at 100 rpm for 24 hours. 2. This culture method: Put 1 ~ 10g of red beans, 0.2g of potassium nitrate, 0.3g of potassium dihydrogen phosphate, and 100ml of φ water into a 500ml Erlenmeyer flask with a plug, and place it in a high-temperature autoclave at 121 ° C. Sterilize for 15 minutes. 1 ml of the pre-culture broth was planted in this medium, and cultured at 37 C with shaking at 1000 r pm for 4 8 hours. After the completion of the culture, the glucoamylase (g A) activity and the acid-resistant α-amylase (A S A A) activity in the culture supernatant were measured in the same manner as in Example 15. 3. Results:

如表34所示。如前述般，釀造燒酒所需的酵素活性的 目標値’葡糖澱粉酶爲l〇〇U/ml、耐酸性α -澱粉酶爲 10U/m卜從表清楚可知，紅豆用量爲2%時GA活性和AS A A -49- 200538545 活性最大，紅豆用量爲1〜2%時，目標酵素活性値變清楚。 由此結果可知，紅豆適合使用爲液體麴之原料。 (表 34 ) 紅丑用量 酵素活性（U/ml) GA ASAA 1% 126.7 11.3 2% 138.2 12.9 4% 85.4 10.8 8% 67.7 9.4 10% 59.0 8.9As shown in Table 34. As mentioned above, the target glucoamylase for enzyme activity required for brewing shochu is 100 U / ml and acid-resistant α-amylase is 10 U / m. It is clear from the table that when the amount of red beans is 2%, GA Activity and AS AA -49- 200538545 have the highest activity. When the amount of red beans is 1 ~ 2%, the target enzyme activity becomes clear. From this result, it can be seen that red beans are suitable for use as a raw material for liquid tincture. (Table 34) Dosage of red and ugly enzyme activity (U / ml) GA ASAA 1% 126.7 11.3 2% 138.2 12.9 4% 85.4 10.8 8% 67.7 9.4 10% 59.0 8.9

(實施例1 8 )以使用紅豆的液體麹製造米燒酒 1.固體麴製造方法 使用90%碾製米，洗米後浸漬15分鐘，瀝乾10分鐘， 並蒸煮30分鐘後使冷卻至40°C，對於每lkg碾製米，植菌 lg 的種麹（白麴菌 Aspergillus kawachii IF0 4308)，於 40 。(：•相對溼度95%的條件培養24小時，又於35 °C ·相對 溼度95%下培養6小時，再於30°C ·相對溼度90%下培養 1 8小時。 2.液體麹之製法 (1 )前培養方法： 8g的90%碾製米和100ml水放入500ml的附栓三角燒 瓶，置於高溫高壓滅菌釜以1 2 1°C進行1 5分鐘滅菌。冷卻 後，將白麴菌（Aspergillus kawachii IF 0 4308)的種麴孢子 植菌於前培養基中，使含量爲lxl 06個/m卜並於37 °C，以 1 〇 〇 r p m震盪培養2 4小時。 (2 )本培養法： -50- 200538545 將10g紅豆和l.Og硝酸鉀、1.5g磷酸二氫鉀和5 00ml 水放入2 0 0 0 m 1的附栓三角燒瓶’置於高溫高壓滅菌釜以 1 2 1 °C進行1 5分鐘滅菌。將5 ml前培養液植菌於此本培養 基，並於37°C，以lOOrpm震盪培養48小時’製造大豆液 體麴。 3 .米燒酒之製法 (1)使用酵母：燒酒酵母（鹿兒島酵母） (2 )釀造掺合： 釀造掺合如表35、表36所示。米係使用90%碾製米， ® 洗米後浸漬1 5分鐘，瀝乾1 0分鐘，並蒸煮3 0分鐘後的米。 試驗區分爲1)固體麴裝入釀造、2)紅豆液體麴裝入釀造 等2試驗區，二試驗區的總米量及水量相等。酵母係將5 0 μ 1於3 0 °C靜置培養4 8小時者植菌於Y P D培養基。 (3 )發酵條件：25°C，固定 (4 )蒸留條件：減壓蒸留 (表 35 ) 試驗區（紅豆液體麹） 第一次 第二次 第三次 合計 麴紅豆（g) 10.0 — _ 10.0 續加入米（g) 311.3 507.6 507.6 1326.5 水（mL ) 594.0 765.4 265.6 1625.0 液體麴（mL) 500.0 一 _ 500.0 90% 乳酸（mL) 1.4 — —— 1.4 -51- 200538545 (表 3 6 ) 對照區（固體麹） 第一次 第二次 第三次 合計 麴米（g) 311.3 — — 311.3 續加入米（g ) 一 507.6 507.6 1015.2 水（m L ) 594.0 765.4 765.6 2125.0 4.結果與硏究 發酵過程如第11圖所示。從圖清楚可知，米固體麴裝入釀造 區和紅豆液體麴裝入釀造區，具有幾乎相同的發酵過程。又，最 φ 後酒醴的酒精度數，米固體麴裝入釀造區19.1% 、紅豆液體麴裝 入釀造區19.2%幾乎相同程度。 以減壓蒸留法使製得固體麴裝入釀造區、紅豆液體麴裝入釀造 區的燒酒酒醴進行蒸留，由6位專業品評員以5點品評法（佳1-3-5 差）進行上述製得的燒酒原酒之官能品評，結果如表37所示，可 知固體麹裝入釀造區和紅豆液體麴裝入釀造區無太大的差異，紅 豆液體麴亦可製造具充分品質的燒酒。又，紅豆液體麴區有「鬆 軟、甘甜香味」之評語，顯示可製造和先前固體麹製法有明顯差 異化之燒酒原酒。 (表 37) 官能品評結果 評點（平均點） 風評 固體麹 3.0 舒暢 紅豆液體麴 2.8 鬆軟、紅豆般甘甜香味 (實施例19)甘藷液體麴之製法 1.前培養方法： 8g的90%碾製米和100ml水放入500ml的附栓三角燒瓶，置 於高溫高壓滅菌釜以121 °C進行15分鐘滅菌。冷卻後，將白麴菌 -52- 200538545 (Aspergillus kawachiiIFO4308 )的種麴孢子植菌於前培養基中， 使含量爲lxlO6個/ml ’並於37°C，以lOOrpm震盪培養24小時。 2.本培養法： 輕柔地清洗1條生甘藷（約20g )的外側，不須進行去蒂或削 皮等處理直接和l.Og硝酸鉀、1.5g磷酸二氫鉀和500ml水放入 2000ml的附栓三角燒瓶，置於高溫高壓滅菌釜以121。(：進行15分 鐘滅菌。將lml前培養液植菌於此本培養基，並於37°C，以80rpm 輕緩地震盪，使培養液中的甘藷不崩解地培養48小時。 培養結束後，以實施例15的方法測定培養上清液中的葡糖澱 粉酶（GA )活性和耐酸性α -澱粉酶（ASAA )活性。 籲3.結果： 如表38所示。如前述般，釀造燒酒所需的酵素活性的目標 値，葡糖澱粉酶爲100U/ml、耐酸性α -澱粉酶爲10U/ml。 從表38清楚可知，可同時生成GA和ASAA二酵素。ASAA 的目標酵素活性値不清楚者，且通氣條件等液體麴培養條件維持 在最適狀況時，期待可增加酵素活性，即使是現狀的甘藷液體麹， 藉著增加燒酒釀造時的麴比率亦可充分地製造燒酒。 (表 38) 甘藷 酵素活性（U/ml) GA ASAA 108.6 5.5 (實施例20 )莧紫液體麹之製法 1.前培養方法：8g的90%碾製米和l〇〇ml水放入500ml 的附栓三角燒瓶，置於高溫高壓滅菌釜以1 2 1 °C進行1 5分 鐘滅菌。冷卻後，將白麴菌（Aspergillus kawachii IF0 4308) 的種麴孢子植菌於前培養基中，使含量爲lxlO6個/ml，並 於3 7 °C ，以1 0 0 r p m震盪培養2 4小時。 -53- 200538545(Example 1 8) Rice shochu made from red rice bean liquid shochu 1. A method for making solid rice vinegar using 90% milled rice, washed rice, soaked for 15 minutes, drained for 10 minutes, and cooked for 30 minutes to cool to 40 ° C For each 1 kg of milled rice, the species of Phytophthora spp. (Aspergillus kawachii IF0 4308), at 40. (: • Cultivate for 24 hours at 95% relative humidity, and then at 35 ° C for 6 hours at 95% relative humidity, and then at 30 ° C for 18 hours at 90% relative humidity. 2. Production method of liquid tincture (1) Pre-cultivation method: 8g of 90% milled rice and 100ml of water are placed in a 500ml Erlenmeyer flask with a plug, and placed in a high-temperature autoclave for sterilization at 1 2 1 ° C for 15 minutes. After cooling, the white tincture (Aspergillus kawachii IF 0 4308) in the pre-culture medium, the content of lxl 06 / mbu and 37 ° C, shaking culture at 100 rpm for 24 hours. (2) the culture Method: -50- 200538545 Put 10 g of red beans and 1.0 g of potassium nitrate, 1.5 g of potassium dihydrogen phosphate and 5,000 ml of water into a 2 0 0 0 m 1 conical Erlenmeyer flask placed in an autoclave to 1 2 1 It is sterilized for 15 minutes at 5 ° C. 5 ml of the pre-culture broth is planted in this medium, and cultured at 37 ° C with shaking at 100 rpm for 48 hours to produce soybean liquid mash. 3. Method for making rice shochu (1) using yeast : Shochu yeast (Kagoshima yeast) (2) Brewing blends: The brewing blends are shown in Table 35 and Table 36. For rice, use 90% milled rice. After washing the rice After soaking for 15 minutes, draining for 10 minutes, and cooking for 30 minutes, the test is divided into 1) solid mashed into brewing, 2) red bean liquid mashed into brewing, etc. 2 test zones, total rice in the second test zone The amount and amount of water are equal. Yeast lines were cultured at 50 μ1 at 30 ° C for 4 to 8 hours, and planted in Y P D medium. (3) Fermentation conditions: 25 ° C, fixed (4) Steam retention conditions: vacuum distillation (Table 35) Test area (red bean liquid tincture) Total amount of red beans (g) 10.0 — _ 10.0 Continue adding Rice (g) 311.3 507.6 507.6 1326.5 Water (mL) 594.0 765.4 265.6 1625.0 Liquid tritium (mL) 500.0 _ 500.0 90% Lactic acid (mL) 1.4 — —— 1.4 -51- 200538545 (Table 3 6) Control area (solid tritium ) Total amount of indica rice (g) for the first, second, and third times 311.3 — — 311.3 Continue to add rice (g)-507.6 507.6 1015.2 Water (m L) 594.0 765.4 765.6 2125.0 4. Results and research fermentation process as shown in Figure 11 Show. It is clear from the figure that the rice solid 麴 loaded into the brewing zone and the red bean liquid 麴 loaded into the brewing zone have almost the same fermentation process. In addition, the alcohol content of the last wine 醴 is almost the same as that of rice solid rice filled 19.1% in the brewing zone and red bean liquid rice packed in the brewing zone 19.2%. The solid 麴 produced in the brewing area and the red bean liquid 麴 into the shochu wine 酿 in the brewing area were steamed and retained by the reduced-pressure steaming method. Six professional judges performed the five-point evaluation method (good 1-3-5 poor). The functional evaluation of the shochu raw liquor prepared as described above is shown in Table 37. It can be seen that there is not much difference between the solid 麹 loading in the brewing zone and the red bean liquid 麴 loading in the brewing zone, and the red bean liquid 麴 can also produce shochu with sufficient quality. In addition, the red bean liquid glutinous area has a comment of "soft and sweet flavor", which shows that it is possible to make shochu original wine that is significantly different from the previous solid mash method. (Table 37) Functional evaluation result evaluation points (average point) Wind evaluation solid 麹 3.0 Shuchang red bean liquid 麴 2.8 Soft, red bean-like sweet flavor (Example 19) Method for making sweet potato liquid 麴 1. Pre-cultivation method: 8g of 90% milled rice Put 100ml of water into a 500ml Erlenmeyer flask with a stopper and place in an autoclave at 121 ° C for 15 minutes. After cooling, the spore spores of Aspergillus kawachiiIFO4308 (Aspergillus kawachiiIFO4308) were planted in the pre-culture medium at a content of 1 × 10 6 cells / ml ′ and cultured at 37 ° C. with shaking at 100 rpm for 24 hours. 2. This culture method: Gently clean the outside of one raw sweet potato (about 20g), and directly put 1.0g potassium nitrate, 1.5g potassium dihydrogen phosphate, and 500ml water into 2000ml without processing the pedicle or peeling. The Erlenmeyer flask with a stopper was placed in an autoclave at 121. (: Sterilize for 15 minutes. Plant 1 ml of the pre-culture broth into this medium, and gently shake at 37 ° C and 80 rpm to culture the sweet potato in the broth without disintegration for 48 hours. After the end of the culture, The glucoamylase (GA) activity and acid-resistant α-amylase (ASAA) activity in the culture supernatant were determined by the method of Example 15. 3. Results: As shown in Table 38. As mentioned above, shochu was brewed The target enzyme activity required for enzyme activity is 100 U / ml for glucoamylase and 10 U / ml for acid-resistant α-amylase. As is clear from Table 38, GA and ASAA enzymes can be produced simultaneously. Target enzyme activity of ASAA If you are not sure, and if the culture conditions of liquid 麴 such as aeration conditions are maintained at optimal conditions, it is expected that the enzyme activity will be increased. Even in the current sweet potato liquid 麹, you can make shochu adequately by increasing the ratio of 麴 during brewing. 38) Enzyme activity of sweet potato (U / ml) GA ASAA 108.6 5.5 (Example 20) Preparation method of 苋 purple liquid 麹 1. Pre-cultivation method: 8 g of 90% milled rice and 100 ml of water were put into 500 ml of suppository Erlenmeyer flask, placed in an autoclave at 1 2 1 ° C Sterilize for 15 minutes. After cooling, the Aspergillus kawachii IF0 4308 spore spores are planted in the pre-culture medium to a content of 1 × 10 6 cells / ml at 37 ° C at 100 rpm. Shake for 24 hours. -53- 200538545

2.本培養法：將1〜8g莧紫和0.2g硝酸鉀、0.3g磷酸二 氫鉀和100ml水放入5 00ml的附栓三角燒瓶，置於高溫高 壓滅菌釜以1 2 1 °C進行1 5分鐘滅菌。將1 ml前培養液植菌 於此本培養基，並於37t，以lOOrpm震盪培養48小時。 培養結束後，測定培養上清液中的葡糖澱粉酶活性（GA ) 和耐酸性α -澱粉酶活性（ASAA )。使用糖化力分別定量 套組（龜甲萬製）測定GA活性，AS ΑΑ活性之測定方法， 係將（非專利文獻4)的方法稍加改良，培養物經過酸處 理使非耐酸性α -澱粉酶失活後，使用α澱粉酶測定套組 (龜甲萬製）進行測定。更具體而言，在1 ml培養液中添 加9ml的100mM醋酸緩衝液（ph3 )，於37°C進行1小時 酸處理後’使用α澱粉酶測定套組（龜甲萬製）進行測定。 3 ·結果：如表3 9所示。至目前爲止的硏究，釀造燒酒所 需的酵素活性的目標値，葡糖澱粉酶爲l〇〇U/nU、耐酸性α •S粉酶爲10U/ml。從表清楚可知，莧紫用量2%以上時， GA、AS AA二酵素的活性的目標値變清楚，顯示莧紫可有 效地作爲液體麴原料。2. This culture method: Put 1 ~ 8g osmanthus violet and 0.2g potassium nitrate, 0.3g potassium dihydrogen phosphate and 100ml water in a 500ml conical Erlenmeyer flask, put it in a high-temperature autoclave at 1 2 1 ° C Sterilize for 15 minutes. 1 ml of the pre-culture broth was planted in this medium, and cultured at 37 t with shaking at 100 rpm for 48 hours. After the culture was completed, the glucoamylase activity (GA) and acid-resistant α-amylase activity (ASAA) in the culture supernatant were measured. The method for measuring GA activity and AS ΑΑ activity using saccharification power respectively quantitative set (made by Kikkoman) is a slightly improved method (non-patent document 4), and the culture is acid-treated to make non-acid-resistant α-amylase. After the inactivation, the measurement was performed using an α-amylase measurement kit (Kikkoman). More specifically, 9 ml of 100 mM acetate buffer (ph3) was added to 1 ml of the culture solution, and the mixture was subjected to acid treatment at 37 ° C for 1 hour. The measurement was performed using an α-amylase measurement kit (made by Kikkoman). 3 Result: as shown in Table 3-9. So far, the target enzyme activity required for brewing shochu is 100 U / nU for glucoamylase and 10 U / ml for acid-resistant α • S powder enzyme. It is clear from the table that when the amount of 苋 purple is more than 2%, the target of the activity of GA and AS AA dienzymes becomes clear, showing that 苋 purple can be effectively used as a liquid 麴 raw material.

(表 39 ) 莧紫用量 卜酵素活， 丨生（U/ml) ___GA ASAA 1% 108.6 5.5 2% 11.0 4% 13.5 8% 13.4 -54- 200538545 (實施例2 1 )奎藜籽液體麹之製法 1 ·前培養方法：8 g的9 0 %碾製米和1 0 0 m 1水放入5 0 0 m 1 的附栓三角燒瓶’置於高溫高壓滅菌釜以1 2 It：進行1 5分 鐘滅囷。冷卻後’將白麴菌（Aspergillus kawachii IF04308) 的種麴孢子植菌於前培養基中，使含量爲1x1 〇6個/m卜並 於3 7 °C，以1 0 0 r p m震盪培養2 4小時。 2 ·本培養法··將1〜8 g的奎藜籽和〇 . 2 g硝酸鉀、0.3 g磷 酸二氫鉀和100ml水放入500ml的附栓三角燒瓶，置於高 溫高壓滅菌釜以1 2 1 °C進行1 5分鐘滅菌。將1 ml前培養液 ® 植菌於此本培養基，並於37°C，以lOOrpm震盪培養48小 時。培養結束後’以實施例20的方法測定培養上清液中的 葡糖澱粉酶活性（GA )和耐酸性α -澱粉酶活性（AS AA )。 3 ·結果：如表4 0所示。至目前爲止的硏究，釀造燒酒所 需的酵素活性的目標値，葡糖澱粉酶爲iOOU/ml、耐酸性α -澱粉酶爲10U/m卜從表清楚可知，奎藜籽用量爲2〜4%時， 可平衡地生成二酵素，顯示奎藜籽可有效地作爲液體麴原料。 (表 40 ) 用量 酵素活性（U/ml) GA ASAA 1% 132.7 7.3 2% 126.8 10.6 4% 123.5 11.1 8% 88.2 18.1 〔應用於產業上的可能性〕 根據本發明’提供一種使用榖皮覆蓋表面之榖類或只去 除表面的榖皮（粗殼）之榖類、外皮覆蓋表面之豆類或芋 -55- 200538545 類、或雜榖類莧紫及/或奎藜籽作爲培養原料，有效地且廉 價地製造品質安定的液體麴之方法。且，此液體麴不僅適 用於發酵飲食品的製造，因可平衡地生成大量的葡糖澱粉 酶及耐酸性α -澱粉酶二酵素，適用於燒酒等酒類之製造。 【圖式簡單說明】 第1圖以使用糙麥的液體培養基進行麴菌培養時，糙麥用 量和葡糖澱粉酶及耐酸性α -澱粉酶的生成量之關係圖。 第2圖以使用對照區的圓麥的液體培養基進行麴菌培養 時，糙麥用量和葡糖澱粉酶及耐酸性澱粉酶的生成量之 關係圖。 第3圖以使用糙麥的液體麴製造麥燒酒時的發酵過程圖。 第4圖以使用喬麥的液體麴製造米燒酒時的發酵過程 圖。 第5圖以使用小米的液體麴製造米燒酒時的發酵過程 圖。 第6圖以使用稗的液體麹製造米燒酒時的發酵過程圖。 第7圖以使用稷的液體麴製造米燒酒時的發酵過程圖。 第8圖以使用高梁的液體麹製造米燒酒時的發酵過程 圖。 第9圖使用各種的麴製造燒酒時的發酵過程圖。 第1 0圖以使用大豆的液體培養基進行麴菌培養’再使 用製得的液體麴製造燒酒時的發酵過程圖。 第1 1圖以使用紅豆的液體培養基進行麴菌培養’再使 用製得的液體麹製造燒酒時的發酵過程圖。 -56 -(Table 39) The amount of pueraria purpurea enzyme activity, health (U / ml) ___GA ASAA 1% 108.6 5.5 2% 11.0 4% 13.5 8% 13.4 -54- 200538545 (Example 2 1) Preparation method of quinoa seed liquid tincture 1 · Pre-cultivation method: 8 g of 90% milled rice and 100 m 1 water into a 500 m 1 conical Erlenmeyer flask placed in an autoclave at 1 2 It: for 15 minutes Extinction. After cooling, the spores of Aspergillus kawachii IF04308 were planted in the pre-culture medium to a content of 1 × 106 cells / m at 37 ° C and shaken at 100 rpm for 2 to 4 hours. . 2 · This culture method · Put 1 ~ 8 g of quinoa seeds and 0.2 g of potassium nitrate, 0.3 g of potassium dihydrogen phosphate and 100 ml of water into a 500 ml Erlenmeyer flask with a suppository. Sterilize at 21 ° C for 15 minutes. 1 ml of pre-culture medium ® was planted in this medium, and cultured at 37 ° C with shaking at 100 rpm for 48 hours. After the cultivation was completed, glucoamylase activity (GA) and acid-resistant α-amylase activity (AS AA) in the culture supernatant were measured by the method of Example 20. 3 Result: as shown in Table 40. So far, the target of enzyme activity required for brewing shochu is glucoamylase iOOU / ml and acid-resistant α-amylase 10U / m. It is clear from the table that the amount of quinoa seeds is 2 ~ At 4%, two enzymes can be produced in a balanced manner, showing that quinoa seeds can be effectively used as a raw material for liquid tincture. (Table 40) Dosage enzyme activity (U / ml) GA ASAA 1% 132.7 7.3 2% 126.8 10.6 4% 123.5 11.1 8% 88.2 18.1 [Possibility of application in the industry] According to the present invention, a surface of the skin is provided by using榖 or 只 only removes the surface of the pupa skin (coarse shell), peas or taro-55-200538545, or mixed pupa purple and / or quinoa seeds as the raw material for cultivation, effectively and A method for inexpensively manufacturing stable liquid plutonium. In addition, this liquid tincture is not only suitable for the production of fermented foods and beverages, but can produce a large amount of glucoamylase and acid-resistant α-amylase dienzyme in a balanced manner, and is suitable for the production of liquor such as shochu. [Brief description of the figure] Fig. 1 is a graph showing the relationship between the amount of brown wheat and the amount of glucoamylase and acid-resistant α-amylase produced when the fungus is cultured in liquid medium using brown wheat. Fig. 2 is a graph showing the relationship between the amount of brown wheat and the amount of glucoamylase and acid amylase production when cultivating the fungi using liquid medium of round wheat in the control area. FIG. 3 is a fermentation process diagram when shochu is produced using liquid mash of brown wheat. Fig. 4 is a diagram showing the fermentation process when rice shochu is produced using Qiaomai's liquid mash. Fig. 5 is a fermentation process diagram when rice shochu is produced using liquid millet of millet. FIG. 6 is a fermentation process diagram when rice shochu is produced using liquid mash of mash. FIG. 7 is a fermentation process diagram when rice shochu is produced using liquid mash of mash. Fig. 8 is a fermentation process diagram of rice shochu using liquid sorghum using sorghum. FIG. 9 is a fermentation process diagram when shochu is produced using various kinds of scallions. Fig. 10 is a diagram showing the fermentation process when shochu culture is carried out using a liquid culture medium of soybeans, and the shochu produced by using the prepared liquid mash. Fig. 11 is a diagram showing the fermentation process when shochu culture is carried out using a liquid medium of red beans, and then the shochu produced by using the prepared liquid mash. -56-

Claims (1)

200538545 十、申請專利範圍： 1.一種液體麴之製法，其特徵係用於製造發酵飲 液體麴之製法，將麴菌培養於含有任一種榖皮覆 之榖類；只去除表面的榖皮（粗殻）之榖類；外 表面的未加工的豆類或芋類；及莧紫及/或奎藜米 作爲培養原料之液體培養基。 2·如申請專利範圍第1項之液體麹之製法，其中 在表面之榖類係未碾製、或以至少有榖皮殘留在 面上以上之碾製比率碾製者。 • 3 ·如申請專利範圍第丨或2項之液體麹之製法， 係大麥。 4 ·如申請專利範圍第3項之液體麴之製法，其中 述碾製比率9 0 %以上者。 5 .如申請專利範圍第1或2項之液體麹之製法， 係米、小麥、喬麥、稗、小米、稷、高梁或玉为 6.如申請專利範圍第1項之液體麴之製法，其中 面的榖皮（粗殼）之榖類係糙米。 φ 7 .如申請專利範圍第1項之液體麹之製法，其中 在表面的未加工的豆類或芋類係大豆、紅豆、驾 8 .如申請專利範圍第1〜7項中任一項之液體麴之 中培養於含上述培養原料的液體培養基之麴_ 中，至少同時生成、蓄積葡糖澱粉酶和耐酸性α 9. 一種發酵飮食品之製法，其特徵係使用以申請 1〜8項中任一項方法製得的液體麴製造發酵飲食 1 0.如申請專利範圍第9項之發酵飮食品之製法， 食品時的 蓋在表面 皮覆蓋在 ? (q u i η 〇 a) 榖皮覆蓋 榖粒的表 其中榖類 大麥係上 其中榖類 S： ° 只去除表 外皮覆蓋 ξ甘藷。 製法，其 菌培養物 -澱粉酶。 專利範圍 品。 其中發酵 -57- 200538545 飲食品的所有製造工程均在液態下進行。 1 1 .如申請專利範圍第9或1 0項之發酵飲食品之製法，其 中發酵飮食品的製造係於與外界保持遮蔽的液態狀態 下進行。 1 2 ·如申請專利範圍第9〜n項中任一項之發酵飲食品之製 法’其中發酵飲食品的製造係由將副原料置入上述液體 麴中，製造一次酒醴而進行。 1 3 ·如申請專利範圍第9〜1 2項中任一項之發酵飲食品之製 法’其中發酵飲食品係燒酒。 1 4 · 一種發酵飲食品製造用的液體麴組，其特徵係至少含有 以申請專利範圍1〜8項中任一項之液體麴之製法製得的 葡糖澱粉酶活性和耐酸性^ —澱粉酶活性。 15·如申請專利範圍第！〜7項中任一項之液體麹之製法，其 中將麴菌培養於含有上述培養原料的液體培養基以製 xa液體麹中’藉著抑制培養原料中由澱粉由來的糖質釋 放至培養基的速度，以調整液體麹的酵素活性。200538545 10. Scope of patent application: 1. A method for manufacturing liquid ravioli, which is characterized by the method for producing fermented drink liquid ravioli, culturing fungi in any kind of coriander containing any kind of coriander; only removing the surface of coriander ( Coarse shells); coriander; raw beans or taro on the outer surface; and liquid culture medium with coriander and / or quinoa as raw materials. 2. The method of manufacturing liquid radon as described in the first item of the patent application scope, wherein the ravioli on the surface is unmilled or is milled at a rolling ratio with at least the skin remaining on the surface. • 3 · If the method of making liquid 麹 in the scope of patent application No. 丨 or 2 is barley. 4 · If the method of making liquid radon in item 3 of the patent application scope, wherein the rolling ratio is more than 90%. 5. If the method of making liquid radon in item 1 or 2 of the scope of patent application is rice, wheat, jojoba, ravioli, millet, ravioli, sorghum or jade. 6. If the method of making liquid radon in item 1 of patent scope, The scallion (coarse shell) of the noodles is brown rice. φ 7. The method for making liquid tincture according to item 1 of the scope of patent application, in which raw beans or taro-based soybeans, red beans, and red beans are used on the surface. 8. The liquid is any one of items 1 to 7 of the scope of patent application麴 is cultured in 麴 _, a liquid medium containing the above-mentioned culture materials, and at least glucoamylase and acid-resistant α are produced and accumulated at the same time. 9. A method for producing fermented 飮 foods, which is characterized by using 1 to 8 applications Liquid fermented food prepared by any of the methods 10. Fermented food made from fermented food prepared according to item 9 of the patent application method, when the food is covered on the surface, the skin is covered with? (Qui η 〇a) The surface of the barley line is in the barley line S: ° Only the outer skin is covered with ξ sweet potato. Preparation method, its bacterial culture-amylase. Scope of patent. Among them fermentation -57- 200538545 All manufacturing processes of food and beverage are carried out in liquid state. 1 1. The method for manufacturing a fermented food or drink product according to item 9 or 10 of the scope of patent application, wherein the manufacturing of fermented glutinous food is carried out in a liquid state kept in the shade from the outside. 1 2 · The method for manufacturing a fermented food or drink according to any one of the items 9 to n of the scope of application for an application ', wherein the production of the fermented food or drink is made by placing the auxiliary raw materials in the above-mentioned liquid mash and manufacturing the wine mash once. 1 3 · The method for producing a fermented food or drink according to any one of the items 9 to 12 of the scope of patent application ', wherein the fermented food or drink is shochu. 1 4 · A liquid mash group for producing fermented food and drink, characterized in that it contains at least the glucoamylase activity and acid resistance ^ prepared by the method of preparing liquid mash of any one of claims 1 to 8 Enzyme activity. 15 · If the scope of patent application is the first! A method for producing liquid mash according to any one of ~ 7, wherein the mash fungus is cultured in a liquid medium containing the above-mentioned culture material to produce xa liquid mash 'by inhibiting the rate of release of starch-derived saccharide from the culture material to the culture medium. To adjust the enzyme activity of liquid puppets. -58--58-
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AU2011200747B2 (en) 2011-12-01

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